Treating ige-mediated allergic diseases

ABSTRACT

Methods for treating a disorder associated with immunoglobulin E (IgE) in a subject with antibodies capable of binding to the Cεmx domain of a membrane-bound IgE. The subject can be administered with at least two doses of the antibody, the two doses being at least three months apart.

RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.16/862,512, filed Apr. 29, 2020, which is a continuation-in-part ofInternational Patent Application No. PCT/CN2018/112714, filed on Oct.30, 2018, which claims the benefit of the filing date under 35 U.S.C. §119 of U.S. Provisional Application Ser. No. 62/579,416, filed Oct. 31,2017, the entire contents of each of which are incorporated by referenceherein.

BACKGROUND OF THE INVENTION

IgE plays a central role in mediating type I hypersensitivity reactionsthat are responsible for causing allergic diseases, including allergicasthma, allergic rhinitis, atopic dermatitis, and others. Allergicreactions are the responses of the immune system toward harmlessenvironmental substances, such as dust mites, tree and grass pollens,certain food and drugs, and bee and fire ant bites. In such reactions,the binding of an allergen to IgE on the surface of basophils and mastcells causes the cross-linking of IgE and the aggregation of theunderlying receptors of IgE.Fc, the type I IgE.Fc receptors, or FcεRI.This receptor aggregation subsequently activates the signaling pathwayleading to the exocytosis of granules and the release of pharmacologicmediators, such as histamine, leukotrienes, tryptase, cytokines andchemokines. The release of those mediators from mast cells and basophilscauses the various pathological manifestations of allergy.

There are two types of IgE molecules, free (or soluble) IgE andmembrane-bound IgE (mIgE). Free IgE molecules circulate in the blood andinterstitial fluid. mIgE are expressed on the surface of B lymphoblastsand memory B cells. Targeting mIgE is believed to be effective ininhibiting the production of antigen-specific IgE and thus suppressingIgE-medicated immune responses.

SUMMARY OF THE INVENTION

The present disclosure is based on the unexpected discovery that asingle dose of FB825, an antibody that targets the CεmX domain of mIgEon human B lymphocytic cells, successfully reduced the level of totalIgE in human subjects for at least three months.

Accordingly, one aspect of the present disclosure provides a method fortreating a disorder associated with IgE, the method comprisingadministering to a subject in need thereof a first dose of an antibodybinding to a Cεmx domain of a membrane-bound IgE; and administering tothe subject a second dose of the antibody. The second dose isadministered at least 8 weeks (e.g., at least 10 weeks, 12 weeks or 3months) after the first dose.

In any of the methods described herein, the first dose, the second dose,or both may range from 0.5 mg/kg to 15 mg/kg (e.g., 1 mg/kg to 15mg/kg). For example, the first dose, the second dose, or both range from1 mg/kg to 8 mg/kg (e.g., 1.5 mg/kg to 10 mg/kg). The first dose, thesecond dose, or both can be administered by intravenous injection.

The subject to be treated by the method described herein may be a humanpatient having or suspected of having the disorder associated with IgE,e.g., allergic asthma, allergic rhinitis, hyper IgE syndrome, or atopicdermatitis. In some embodiments, the disorder is cold-induced urticaria,chronic urticaria, cholinergic urticaria, chronic rhinosinusitis,systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonaryaspergillosis, recurrent idiopathic angioedema, and interstitialcystitis, eosinophil-associated gastrointestinal disorders, a foodallergy, or a drug allergy.

In another aspect, the present disclosure provides a method of treatingatopic dermatitis, the method comprising administering to a subject inneed thereof a first dose of an antibody binding to a Cεmx domain of amembrane-bound IgE; wherein the first dose is about 1 mg/kg to 10 mg/kg(e.g., 3 mg/kg to 8 mg/kg, e.g., 5 mg/kg). The method may furthercomprise administering to the subject a second dose of the antibodyabout 3 months after the first dose, if at the time of the second dose,the change of the total IgE level in the subject from the total IgElevel before the first dose is less than 50%. In some instances, thesecond dose may be identical to the first dose, e.g., 5 mg/kg. The firstdose of the antibody, the second dose of the antibody, or both may beadministered by intravenous infusion.

In any of the method described above, a moisturizer is applied to thesubject at least twice a day for at least seven consecutive days priorto the first dose. Alternatively or in addition, the method may furthercomprise administering to the subject a topical corticosteroid. In someinstances, the topical corticosteroid is applied to an active lesiondaily. Such a topical corticosteroid may be a 0.05% fluticasonepropionate cream, a 0.1% monetasone furoate cream, a 0.06% betamethasonevalerate, or a 1% hydrocortisone ointment. In other instances, thetopical corticosteroid may be a 0.05% fluocinonide cream, a 0.25%desoximetasone ointment, or a 0.05% clobetasol propionate ointment.

In some embodiments, the subject is free of topical tacrolimustreatment, topical pimecrolimus treatment, systemic corticosteroidtreatment, leukotriene inhibitor treatment, allergen immunotherapy, atreatment involving an immunosuppressive or immunomodulating agent,vaccine treatment, a treatment involving a traditional Chinese medicine,a surgical procedure, an ultraviolet procedure, or tanning.

In any of the treatment methods described herein, the anti-CεmX antibodydescribed herein may bind the mIgE fragment GLAGGSAQSQRAPDRVL (SEQ IDNO:1) or the mIgE fragment GLAGGSAQSQRA (SEQ ID NO:7). In someinstances, the antibody binds to the same epitope as antibody 4B12(FB825) or competes against antibody FB825 from binding to the Cεmxdomain of a membrane-bound IgE. In some examples, the antibody comprisesthe same heavy chain complementary determining regions as antibody FB825and/or the same light chain complementary determining regions asantibody FB825. Such an antibody may be a humanized antibody of 4B12,for example, FB825. The antibody may comprise a heavy chain variableregion having the amino acid sequence of SEQ ID NO: 2, SEQ ID NO:8, orSEQ ID NO:9; and/or a light chain variable region having the amino acidsequence of SEQ ID NO: 3 or SEQ ID NO:10. Any of the antibodies used inany of the methods described herein can be full-length antibody or anantigen-binding fragment thereof. The antibody can be a human antibody,a humanized antibody, a chimeric antibody, or a single-chain antibody.In some embodiments, the antibody can be an IgG1 molecule. In specificexamples, the antibody may comprise a heavy chain of SEQ ID NO:11 and alight chain of SEQ ID NO:12 (with or without the signal peptide in bothsequences).

In any of the methods described herein, the anti-CεmX antibody can beformulated in a pharmaceutical composition comprising the antibody, abuffer (e.g., a buffer comprising an amino acid such as histidine), asalt (e.g., sodium chloride), and a nonionic surfactant (e.g.,polysorbate 80). In some embodiments, the pharmaceutical composition isan aqueous solution having a pH of 5 to 8. In some examples, theantibody in the pharmaceutical composition is about 10 mg/ml to 30 mg/ml(e.g., about 20 mg/ml), the histidine buffer is of a concentration ofabout 10-30 mM (e.g., about 20 mM), the sodium chloride is of aconcentration of about 120-160 mM (e.g., about 140 mM), and thepolysorbate 80 is of a concentration of about 0.01-0.03% (e.g., about0.02%). Any of the pharmaceutical compositions described herein is alsowithin the scope of the present disclosure.

In another aspect, provided herein is an aqueous formulation, comprisingany of the anti-CεmX antibody described herein (e.g., FB825 or afunctional variant thereof) at a concentration about 10 mg/ml to 30mg/ml, a buffer comprising an amino acid (e.g., histidine) at aconcentration of about 10-30 mM, a salt (e.g., sodium chloride) at aconcentration of about 120-160 mM, and a nonionic surfactant (e.g.,polysorbate 80) at a concentration of about 0.01-0.03%, wherein theaqueous formulation has a pH of about 5-8. In one example, the aqueousformulation comprises the antibody is at a concentration of about 20mg/ml, the histidine buffer is at a concentration of about 20 mM, thesodium chloride is at a concentration of about 140 mM, and thepolysorbate 80 is at a concentration of about 0.02%, and wherein theaqueous formulation has a pH of about 6.5.

Also within the scope of the present disclosure are (i) a pharmaceuticalcomposition for use in treating IgE-associated disorders as describedherein, wherein the pharmaceutical composition comprises an anti-CεmXantibody and a pharmaceutically acceptable carrier, and wherein thepharmaceutical composition is administered to a subject in need of thetreatment for at least two doses, which are at least 8 weeks (e.g., atleast 10 weeks, 12 weeks, or three months apart, or 12 weeks to 6 monthsapart); and (ii) uses of the anti-CεmX antibody in manufacturing amedicament for use in treating the IgE-associated disorder, wherein themedicament can be administered to a subject in need of the treatment forat least two doses, which are at least three months apart.

The details of one or more embodiments of the invention are set forth inthe description below. Other features or advantages of the presentinvention will be apparent from the following drawings and detaileddescription of several embodiments, and also form the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram showing the mean (±SD) change from baseline in totalIgE over time in human subjects treated by a single dose of FB825. TotalIgE was determined from blood samples. Baseline was defined as the lastnon-missing assessment (including repeated and unscheduled assessments)before study drug administration.

FIG. 2 is a diagram showing the mean (±SD) percent change from baselinein total IgE over time in human subjects treated by a single dose ofFB825. Total IgE levels in blood samples obtained from the humansubjects were determined. Baseline was defined as the last non-missingassessment (including repeated and unscheduled assessments) before studydrug administration.

FIG. 3 is a diagram showing the mean (±SD) change from baseline inantidrug antibody (ADA) over time in human subjects treated by a singledose of FB825. ADA in blood samples obtained from the subjects wasdetermined. Baseline is defined as the last non-missing assessment(including repeated and unscheduled assessments) before the study drugadministration.

FIG. 4 shows the amino acid sequence alignment of the V_(H) and V_(L) ofmonoclonal antibody 4B12 and monoclonal antibody FB825, which is ahumanized 4B12 antibody. Variations between the two antibodies arehighlighted. The V_(H) and V_(L) complementary determining regions(CDRs) are in boldface and underlined. V_(H) of 4B12: SEQ ID NO:4. V_(L)of 4B12: SEQ ID NO:5. V_(H) of FB825: SEQ ID NO:2. V_(L) of FB825: SEQID NO:3.

FIG. 5 is a diagram showing the percentage change from baseline inEczema Area and Severity Index (EASI) at Days 8, 15, 29, 57, 85, 92, 99,113, 141, and 169 in the Open-LabeleExploratory Study to Evaluate Safetyand Efficacy of FB825 in Adults with Atopic Dermatitis.

FIG. 6 is a diagram showing the percentage change from baseline inInvestigator's Global Assessment (IGA) at Days 8, 15, 29, 57, 85, 92,99, 113, 141, and 169 in the Open-LabeleExploratory Study to EvaluateSafety and Efficacy of FB825 in Adults with Atopic Dermatitis.

FIG. 7 is a diagram showing the percentage change from baseline inSeverity Scoring of Atopic Dermatitis Index (SCORAD) at Days 8, 15, 29,57, 85, 92, 99, 113, 141, and 169 in the Open-LabeleExploratory Study toEvaluate Safety and Efficacy of FB825 in Adults with Atopic Dermatitis.

FIG. 8 is a diagram showing the percentage change from baseline inPruritus Visual Analogue Scale (VAS) at Days 8, 15, 29, 57, 85, 92, 99,113, 141, and 169 in the Open-LabeleExploratory Study to Evaluate Safetyand Efficacy of FB825 in Adults with Atopic Dermatitis.

DETAILED DESCRIPTION OF THE INVENTION

In atopic individuals who are at increased risk of developing allergies,the IgE concentration in the circulatory system may reach over 10 timesthe normal level. The concentration of allergen-specific IgE antibody isclosely correlated with clinical symptoms and may be over 1000 timeshigher in patients with allergic diseases than in healthy individuals.Immunoglobulin E sensitizes effector cells such as basophils, mastcells, and activated eosinophils by occupying the high-affinity IgEreceptor, FcεRI, on which they are expressed. In type Ihypersensitivity, allergens cross-link IgE molecules bound by FcεRI andsubsequently trigger the degranulation of effector cells, releasingproinflammatory mediators, such as histamines and leukotrienes. TheIgE-mediated allergic pathway, which generates mediator-related allergicsymptoms, initiates immune activities locally or systemically. Basophilsand mast cells also release a wide spectrum of inflammatory cytokinesand chemokines that not only cause clinical symptoms directly but alsoactivate and recruit various cell types to augment inflammatory status.Hence, anti-IgE therapy can attenuate both the IgE-mediated pathway andinflammatory conditions.

Described herein are anti-CεmX antibodies for use in reducing the totalIgE level and thus treating Ig-E medicated disorders. Such antibodiescan be administered to a subject in need of the treatment by at leasttwo doses, which can be at least 8 weeks (e.g., 10 weeks, 12 weeks or 3months) apart.

Antibodies Capable of Binding to a CεmX Domain of a Membrane-Bound IgE

CεmX is a 52-amino acid segment located between the CH4 domain and theC-terminal membrane-anchoring segment of human membrane-bound ε chain(mε). The amino acid sequence of an exemplary CεmX fragment of humanmIgE is provided below (SEQ ID NO:6):

GLAGGSAQSQ RAPDRVLCHS GQQQGLPRAA GGSVPHPRCH CGAGRADWPG PP

The antibodies described herein can bind to the CεmX domain of a mIgE,for example, mIgE expressed on the surface of B cells. Such antibodiesmay induce cell death of the B cells expressing mIgE via, for example,antibody-dependent cell cytotoxicity and/or cell apoptosis, therebyeliminate the B cells, which would lead to reduced production of freeIgE. Accordingly, the anti-CεmX antibodies described herein can reducethe level of total IgE in a subject (e.g., a human patient) beingtreated with the antibody.

An antibody (interchangeably used in plural form) is an immunoglobulinmolecule capable of specific binding to a target, such as acarbohydrate, polynucleotide, lipid, polypeptide, etc., through at leastone antigen recognition site, located in the variable region of theimmunoglobulin molecule. As used herein, the term “antibody” encompassesnot only intact (i.e., full-length) polyclonal or monoclonal antibodies,but also antigen-binding fragments thereof (such as Fab, Fab′, F(ab′)₂,Fv), single chain (scFv), mutants thereof, fusion proteins comprising anantibody portion, humanized antibodies, chimeric antibodies, diabodies,linear antibodies, single chain antibodies, multispecific antibodies(e.g., bispecific antibodies) and any other modified configuration ofthe immunoglobulin molecule that comprises an antigen recognition siteof the required specificity, including glycosylation variants ofantibodies, amino acid sequence variants of antibodies, and covalentlymodified antibodies. An antibody includes an antibody of any class, suchas IgD, IgE, IgG, IgA, or IgM (or sub-class thereof), and the antibodyneed not be of any particular class. Depending on the antibody aminoacid sequence of the constant domain of its heavy chains,immunoglobulins can be assigned to different classes. There are fivemajor classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, andseveral of these may be further divided into subclasses (isotypes),e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy-chain constantdomains that correspond to the different classes of immunoglobulins arecalled alpha, delta, epsilon, gamma, and mu, respectively. The subunitstructures and three-dimensional configurations of different classes ofimmunoglobulins are well known.

The antibodies to be used in the methods described herein can be murine,rat, human, or any other origin (including chimeric or humanizedantibodies).

Any of the antibodies described herein can be either monoclonal orpolyclonal. A “monoclonal antibody” refers to a homogenous antibodypopulation and a “polyclonal antibody” refers to a heterogenous antibodypopulation. These two terms do not limit the source of an antibody orthe manner in which it is made.

In one example, the antibody used in the methods described herein is ahumanized antibody. Humanized antibodies refer to forms of non-human(e.g. murine) antibodies that are specific chimeric immunoglobulins,immunoglobulin chains, or antigen-binding fragments thereof that containminimal sequence derived from non-human immunoglobulin. For the mostpart, humanized antibodies are human immunoglobulins (recipientantibody) in which residues from a complementary determining region(CDR) of the recipient are replaced by residues from a CDR of anon-human species (donor antibody) such as mouse, rat, or rabbit havingthe desired specificity, affinity, and capacity. In some instances, Fvframework region (FR) residues of the human immunoglobulin are replacedby corresponding non-human residues. Furthermore, the humanized antibodymay comprise residues that are found neither in the recipient antibodynor in the imported CDR or framework sequences, but are included tofurther refine and optimize antibody performance. In general, thehumanized antibody will comprise substantially all of at least one, andtypically two, variable domains, in which all or substantially all ofthe CDR regions correspond to those of a non-human immunoglobulin andall or substantially all of the FR regions are those of a humanimmunoglobulin consensus sequence. The humanized antibody optimally alsowill comprise at least a portion of an immunoglobulin constant region ordomain (Fc), typically that of a human immunoglobulin. Antibodies mayhave Fc regions modified as described in WO 99/58572. Other forms ofhumanized antibodies have one or more CDRs (one, two, three, four, five,six) which are altered with respect to the original antibody, which arealso termed one or more CDRs “derived from” one or more CDRs from theoriginal antibody. Humanized antibodies may also involve affinitymaturation.

In another example, the antibody described herein is a chimericantibody, which can include a heavy constant region and a light constantregion from a human antibody. Chimeric antibodies refer to antibodieshaving a variable region or part of variable region from a first speciesand a constant region from a second species. Typically, in thesechimeric antibodies, the variable region of both light and heavy chainsmimics the variable regions of antibodies derived from one species ofmammals (e.g., a non-human mammal such as mouse, rabbit, and rat), whilethe constant portions are homologous to the sequences in antibodiesderived from another mammal such as human. In some embodiments, aminoacid modifications can be made in the variable region and/or theconstant region.

In some examples, the antibody disclosed herein specifically binds aCεmx domain of a membrane-bound IgE, which may be expressed on thesurface of a B cell. An antibody that “specifically binds” (usedinterchangeably herein) to a target or an epitope is a term wellunderstood in the art, and methods to determine such specific bindingare also well known in the art. A molecule is said to exhibit “specificbinding” if it reacts or associates more frequently, more rapidly, withgreater duration and/or with greater affinity with a particular targetantigen than it does with alternative targets. An antibody “specificallybinds” to a target antigen if it binds with greater affinity, avidity,more readily, and/or with greater duration than it binds to othersubstances. For example, an antibody that specifically (orpreferentially) binds to a CεmX domain epitope is an antibody that bindsthis CεmX domain epitope with greater affinity, avidity, more readily,and/or with greater duration than it binds to other CεmX domain epitopesor non-CεmX domain epitopes. It is also understood by reading thisdefinition that, for example, an antibody that specifically binds to afirst target antigen may or may not specifically or preferentially bindto a second target antigen. As such, “specific binding” or “preferentialbinding” does not necessarily require (although it can include)exclusive binding. Generally, but not necessarily, reference to bindingmeans preferential binding.

The binding affinity of an anti-CεmX antibody described herein can beless than about 100 nM, e.g., less than about 50 nM, about 10 nM, about1 nM, about 500 pM, about 100 pM, or about 50 pM to any of about 2 pM.Binding affinity can be expressed K_(D) or dissociation constant, and anincreased binding affinity corresponds to a decreased K_(D). One way ofdetermining binding affinity of antibodies to CεmX is by measuringbinding affinity of monofunctional Fab fragments of the antibody. Toobtain monofunctional Fab fragments, an antibody (for example, IgG) canbe cleaved with papain or expressed recombinantly. The affinity of ananti-CεmX Fab fragment of an antibody can be determined by surfaceplasmon resonance (BIAcore3000™ surface plasmon resonance (SPR) system,BIAcore, INC, Piscaway N.J.). Kinetic association rates (k_(on)) anddissociation rates (k_(off)) (generally measured at 25° C.) areobtained; and equilibrium dissociation constant (K_(D)) values arecalculated as koff/kon.

In some embodiments, the antibody binds the CεmX domain of a human IgE,and does not significantly bind an IgE from another mammalian species.In some embodiments, the antibody binds human IgE as well as one or moreIgE from another mammalian species. The epitope(s) bound by the antibodycan be continuous or discontinuous.

In some embodiments, the anti-CεmX antibody described herein binds anN-terminal portion of the CεmX domain, e.g., GLAGGSAQSQRAPDRVL (SEQ IDNO:1) or GLAGGSAQSQRA (SEQ ID NO:7). Such an antibody may have the sameheavy chain and/or light chain CDRs as antibody 4B12/FB825 as describedin FIG. 4 . See also U.S. Pat. No. 8,460,664, the relevant disclosurestherein are incorporated by reference herein. The anti-CεmX antibody maybe a humanized antibody of 4B12 (e.g., FB825). In some examples, theanti-CεmX antibody for use in the methods described herein is FB825,which is a humanized antibody of 4B12 (FIG. 4 ), or a functional variantthereof. See also U.S. Pat. No. 8,460,664, the relevant disclosurestherein are incorporated by reference herein.

Two antibodies having the same V_(H) and/or V_(L) CDRs means that theirCDRs are identical when determined by the same approach (e.g., the Kabatapproach, the Chothia approach, the AbM approach, the Contact approach,or the IMGT approach as known in the art. See, e.g.,bioinf.org.uk/abs/).

A functional variant (equivalent) of FB825 has essentially the sameepitope-binding specificity as FB825 and exhibits substantially similarbioactivity as FB825, including the activity of eliminating B cellsexpressing mIgE and reducing the level of total IgE in a subject. Insome embodiments, a functional variant of FB825 contains the sameregions/residues responsible for antigen-binding as FB825, such as thesame specificity-determining residues in the CDRs or the whole CDRs. Inother embodiments, a functional variant of FB825 comprises a V_(H) chainthat includes a V_(H) CDR1, V_(H) CDR2, and V_(H) CDR3 at least 75%(e.g., 80%, 85%, 90%, 95%, or 98%) identical to the corresponding V_(H)CDRs of FB825, and a V_(L) chain that includes a V_(L) CDR1, V_(L) CDR2,and V_(L) CDR3 at least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identicalto the corresponding V_(H) CDRs of FB825. For example, a functionalvariant of FB825 may comprise a V_(H) chain that includes up to 5 (e.g.,1, 2, 3, 4, or 5) amino acid residue variations in the V_(H) CDR regions(V_(H) CDR1, CDR2, and/or CDR3 in total) as compared to the V_(H) CDRsof mAb7E, and/or a V_(L) chain that includes up to 5 (e.g., 1, 2, 3, 4,or 5) amino acid residue variations in the V_(L) CDR regions (V_(L)CDR1, CDR2, and/or CDR3 in total) as compared to the V_(H) CDRs ofmAb7E.

Alternatively, the functional variant of FB825 comprises a V_(H) chainat least 75% (e.g., 80%, 85%, 90%, 95%, or 98%) identical to the V_(H)chain of FB825 and a V_(L) chain at least 75% (e.g., 80%, 85%, 90%, 95%,or 98%) identical to the V_(L) chain of FB825. The amino acid sequencevariations may occur only in one or more of the V_(H) and/or V_(L)framework regions.

The “percent identity” of two amino acid sequences is determined usingthe algorithm of Karlin and Altschul Proc. Natl. Acad. Sci. USA87:2264-68, 1990, modified as in Karlin and Altschul Proc. Natl. Acad.Sci. USA 90:5873-77, 1993. Such an algorithm is incorporated into theNBLAST and XBLAST programs (version 2.0) of Altschul, et al. J. Mol.Biol. 215:403-10, 1990. BLAST protein searches can be performed with theXBLAST program, score=50, wordlength=3 to obtain amino acid sequenceshomologous to the protein molecules of interest. Where gaps existbetween two sequences, Gapped BLAST can be utilized as described inAltschul et al., Nucleic Acids Res. 25(17):3389-3402, 1997. Whenutilizing BLAST and Gapped BLAST programs, the default parameters of therespective programs (e.g., XBLAST and NBLAST) can be used.

In some embodiments, the anti-CεmX antibody for use in the treatmentmethod disclosed herein may have one of the following heavy chainvariable regions (CDRs following the Kabat definition are in boldfaceand underlined):

(SEQ ID NO: 2) QVQLQESGPGLVKPSETLSLTCTVS GYSITSDYAWN WIRQPPGKGLEWIGSISYSGITGYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYCA RMG YDGLAY WGQGTLVTVSS(SEQ ID NO: 8) QVQLQESGPGLVKPSETLSLTCTVS GYSITSDYAWN WIRQPPGKGLEWMISISYSGITGYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYCA RMG YDGLAY WGQGTLVTVSS(SEQ ID NO: 9) QVQLQESGPGLVKPSETLSLTCTVS GYSITSDYAWN WIRQPPGKGLEWIGSISYSGITGYNPSLKS RVTISRDTSKNQFSLKLSSVTAADTAVYYCA RMG YDGLAY WGQGTLVTVSS

Alternatively or in addition, the anti-CεmX antibody for use in thetreatment method disclosed herein may have one of the following lightchain variable regions (CDRs following the Kabat definition are inboldface and underlined):

(SEQ ID NO: 3) DIVMTQTPLSLSVTPGQPASISC RSSQSIVHSNGNTYLE WYLQKPGQSPQ LLIYKVSNRFS GVPDRFSGSGSGTEFTLKISRVEAEDVGVYYC FQGSHVP PT FGGGTKVEIKR(SEQ ID NO: 10) DIVMTQTPLSLSVTPGQPASISC RSSQSIVHSNGNTYLE WYLQKPGQSPQLLIY KVSNRFS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC FQGSHVP PT FGGGTKVEIKR

Antibody Preparation

Antibodies capable of binding a CεmX domain of a membrane-bound IgE asdescribed herein can be made by any method known in the art. See, forexample, Harlow and Lane, (1988) Antibodies: A Laboratory Manual, ColdSpring Harbor Laboratory, New York.

In some embodiments, antibodies specific to a target antigen (e.g., aCεmX domain of a mIgE such as a human mIgE) can be made by theconventional hybridoma technology. The full-length target antigen or afragment thereof, optionally coupled to a carrier protein such as KLH,can be used to immunize a host animal for generating antibodies bindingto that antigen. The route and schedule of immunization of the hostanimal are generally in keeping with established and conventionaltechniques for antibody stimulation and production, as further describedherein. General techniques for production of mouse, humanized, and humanantibodies are known in the art and are described herein. It iscontemplated that any mammalian subject including humans or antibodyproducing cells therefrom can be manipulated to serve as the basis forproduction of mammalian, including human hybridoma cell lines.Typically, the host animal is inoculated intraperitoneally,intramuscularly, orally, subcutaneously, intraplantar, and/orintradermally with an amount of immunogen, including as describedherein.

Hybridomas can be prepared from the lymphocytes and immortalized myelomacells using the general somatic cell hybridization technique of Kohler,B. and Milstein, C. (1975) Nature 256:495-497 or as modified by Buck, D.W., et al., In Vitro, 18:377-381 (1982). Available myeloma lines,including but not limited to X63-Ag8.653 and those from the SalkInstitute, Cell Distribution Center, San Diego, Calif., USA, may be usedin the hybridization. Generally, the technique involves fusing myelomacells and lymphoid cells using a fusogen such as polyethylene glycol, orby electrical means well known to those skilled in the art. After thefusion, the cells are separated from the fusion medium and grown in aselective growth medium, such as hypoxanthine-aminopterin-thymidine(HAT) medium, to eliminate unhybridized parent cells. Any of the mediadescribed herein, supplemented with or without serum, can be used forculturing hybridomas that secrete monoclonal antibodies. As anotheralternative to the cell fusion technique, EBV immortalized B cells maybe used to produce the anti-CεmX monoclonal antibodies of the subjectinvention. The hybridomas are expanded and subcloned, if desired, andsupernatants are assayed for anti-immunogen activity by conventionalimmunoassay procedures (e.g., radioimmunoassay, enzyme immunoassay, orfluorescence immunoassay).

Hybridomas that may be used as source of antibodies encompass allderivatives, progeny cells of the parent hybridomas that producemonoclonal antibodies capable of binding the CεmX domain. Hybridomasthat produce such antibodies may be grown in vitro or in vivo usingknown procedures. The monoclonal antibodies may be isolated from theculture media or body fluids, by conventional immunoglobulinpurification procedures such as ammonium sulfate precipitation, gelelectrophoresis, dialysis, chromatography, and ultrafiltration, ifdesired. Undesired activity if present, can be removed, for example, byrunning the preparation over adsorbents made of the immunogen attachedto a solid phase and eluting or releasing the desired antibodies off theimmunogen. Immunization of a host animal with a target antigen or afragment containing the target amino acid sequence conjugated to aprotein that is immunogenic in the species to be immunized, e.g.,keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, orsoybean trypsin inhibitor using a bifunctional or derivatizing agent,for example maleimidobenzoyl sulfosuccinimide ester (conjugation throughcysteine residues), N-hydroxysuccinimide (through lysine residues),glutaraldehyde, succinic anhydride, SOCl, or R1N═C═NR, where R and R1are different alkyl groups, can yield a population of antibodies (e.g.,monoclonal antibodies).

If desired, an antibody (monoclonal or polyclonal) of interest (e.g.,produced by a hybridoma) may be sequenced and the polynucleotidesequence may then be cloned into a vector for expression or propagation.The sequence encoding the antibody of interest may be maintained invector in a host cell and the host cell can then be expanded and frozenfor future use. In an alternative, the polynucleotide sequence may beused for genetic manipulation to “humanize” the antibody or to improvethe affinity (affinity maturation), or other characteristics of theantibody. For example, the constant region may be engineered to moreresemble human constant regions to avoid immune response if the antibodyis used in clinical trials and treatments in humans. It may be desirableto genetically manipulate the antibody sequence to obtain greateraffinity to the target antigen and greater efficacy in reducing totalIgE. It will be apparent to one of skill in the art that one or morepolynucleotide changes can be made to the antibody and still maintainits binding specificity to the target antigen.

In other embodiments, fully human antibodies can be obtained by usingcommercially available mice that have been engineered to expressspecific human immunoglobulin proteins. Transgenic animals that aredesigned to produce a more desirable (e.g., fully human antibodies) ormore robust immune response may also be used for generation of humanizedor human antibodies. Examples of such technology are Xenomouse® fromAmgen, Inc. (Fremont, Calif.) and HuMAb-Mouse® and TC Mouse™ fromMedarex, Inc. (Princeton, N.J.). In another alternative, antibodies maybe made recombinantly by phage display technology. See, for example,U.S. Pat. Nos. 5,565,332; 5,580,717; 5,733,743; and 6,265,150; andWinter et al., (1994) Annu. Rev. Immunol. 12:433-455. Alternatively, thephage display technology (McCafferty et al., (1990) Nature 348:552-553)can be used to produce human antibodies and antibody fragments in vitro,from immunoglobulin variable (V) domain gene repertoires fromunimmunized donors.

Antigen-binding fragments of an intact antibody (full-length antibody)can be prepared via routine methods. For example, F(ab′)2 fragments canbe produced by pepsin digestion of an antibody molecule, and Fabfragments that can be generated by reducing the disulfide bridges ofF(ab′)2 fragments.

Genetically engineered antibodies, such as humanized antibodies,chimeric antibodies, single-chain antibodies, and bi-specificantibodies, can be produced via, e.g., conventional recombinanttechnology. In one example, DNA encoding a monoclonal antibodiesspecific to a target antigen can be readily isolated and sequenced usingconventional procedures (e.g., by using oligonucleotide probes that arecapable of binding specifically to genes encoding the heavy and lightchains of the monoclonal antibodies). The hybridoma cells serve as apreferred source of such DNA. Once isolated, the DNA may be placed intoone or more expression vectors, which are then transfected into hostcells such as E. coli cells, simian COS cells, Chinese hamster ovary(CHO) cells, or myeloma cells that do not otherwise produceimmunoglobulin protein, to obtain the synthesis of monoclonal antibodiesin the recombinant host cells. See, e.g., PCT Publication No. WO87/04462. The DNA can then be modified, for example, by substituting thecoding sequence for human heavy and light chain constant domains inplace of the homologous murine sequences, Morrison et al., (1984) Proc.Nat. Acad. Sci. 81:6851, or by covalently joining to the immunoglobulincoding sequence all or part of the coding sequence for anon-immunoglobulin polypeptide. In that manner, genetically engineeredantibodies, such as “chimeric” or “hybrid” antibodies; can be preparedthat have the binding specificity of a target antigen.

Techniques developed for the production of “chimeric antibodies” arewell known in the art. See, e.g., Morrison et al. (1984) Proc. Natl.Acad. Sci. USA 81, 6851; Neuberger et al. (1984) Nature 312, 604; andTakeda et al. (1984) Nature 314:452.

Methods for constructing humanized antibodies are also well known in theart. See, e.g., Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033(1989). In one example, variable regions of V_(H) and V_(L) of a parentnon-human antibody are subjected to three-dimensional molecular modelinganalysis following methods known in the art. Next, framework amino acidresidues predicted to be important for the formation of the correct CDRstructures are identified using the same molecular modeling analysis. Inparallel, human V_(H) and V_(L) chains having amino acid sequences thatare homologous to those of the parent non-human antibody are identifiedfrom any antibody gene database using the parent V_(H) and V_(L)sequences as search queries. Human V_(H) and V_(L) acceptor genes arethen selected.

The CDR regions within the selected human acceptor genes can be replacedwith the CDR regions from the parent non-human antibody or functionalvariants thereof. When necessary, residues within the framework regionsof the parent chain that are predicted to be important in interactingwith the CDR regions (see above description) can be used to substitutefor the corresponding residues in the human acceptor genes.

A single-chain antibody can be prepared via recombinant technology bylinking a nucleotide sequence coding for a heavy chain variable regionand a nucleotide sequence coding for a light chain variable region.Preferably, a flexible linker is incorporated between the two variableregions. Alternatively, techniques described for the production ofsingle chain antibodies (U.S. Pat. Nos. 4,946,778 and 4,704,692) can beadapted to produce a phage scFv library and scFv clones specific to IgEcan be identified from the library following routine procedures.

Antibodies obtained following a method known in the art and describedherein can be characterized using methods well known in the art. Forexample, one method is to identify the epitope to which the antigenbinds, or “epitope mapping.” There are many methods known in the art formapping and characterizing the location of epitopes on proteins,including solving the crystal structure of an antibody-antigen complex,competition assays, gene fragment expression assays, and syntheticpeptide-based assays, as described, for example, in Chapter 11 of Harlowand Lane, Using Antibodies, a Laboratory Manual, Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y., 1999. In an additionalexample, epitope mapping can be used to determine the sequence to whichan antibody binds. The epitope can be a linear epitope, i.e., containedin a single stretch of amino acids, or a conformational epitope formedby a three-dimensional interaction of amino acids that may notnecessarily be contained in a single stretch (primary structure linearsequence). Peptides of varying lengths (e.g., at least 4-6 amino acidslong) can be isolated or synthesized (e.g., recombinantly) and used forbinding assays with an antibody. In another example, the epitope towhich the antibody binds can be determined in a systematic screening byusing overlapping peptides derived from the target antigen sequence anddetermining binding by the antibody. According to the gene fragmentexpression assays, the open reading frame encoding the target antigen isfragmented either randomly or by specific genetic constructions and thereactivity of the expressed fragments of the antigen with the antibodyto be tested is determined. The gene fragments may, for example, beproduced by PCR and then transcribed and translated into protein invitro, in the presence of radioactive amino acids. The binding of theantibody to the radioactively labeled antigen fragments is thendetermined by immunoprecipitation and gel electrophoresis. Certainepitopes can also be identified by using large libraries of randompeptide sequences displayed on the surface of phage particles (phagelibraries). Alternatively, a defined library of overlapping peptidefragments can be tested for binding to the test antibody in simplebinding assays. In an additional example, mutagenesis of an antigenbinding domain, domain swapping experiments and alanine scanningmutagenesis can be performed to identify residues required, sufficient,and/or necessary for epitope binding. For example, domain swappingexperiments can be performed using a mutant of a target antigen in whichvarious fragments of the IgE polypeptide have been replaced (swapped)with sequences from a closely related, but antigenically distinctprotein (such as another member of the immunoglobulin protein family).By assessing binding of the antibody to the mutant immunoglobulin, theimportance of the particular antigen fragment to antibody binding can beassessed.

Alternatively, competition assays can be performed using otherantibodies known to bind to the same antigen to determine whether anantibody binds to the same epitope as the other antibodies. Competitionassays are well known to those of skill in the art.

Pharmaceutical Compositions

One or more of the above-described anti-CεmX antibodies can be mixedwith a pharmaceutically acceptable carrier (excipient), includingbuffer, to form a pharmaceutical composition for use in treating adisorder associated with IgE. “Acceptable” means that the carrier mustbe compatible with the active ingredient of the composition (andpreferably, capable of stabilizing the active ingredient) and notdeleterious to the subject to be treated. Pharmaceutically acceptableexcipients (carriers) including buffers, which are well known in theart. See, e.g., Remington: The Science and Practice of Pharmacy 20th Ed.(2000) Lippincott Williams and Wilkins, Ed. K. E. Hoover. In oneexample, a pharmaceutical composition contains more than one anti-CεmXantibodies that recognize different epitopes of the target antigen.

The pharmaceutical compositions to be used in the present methods cancomprise pharmaceutically acceptable carriers, excipients, orstabilizers in the form of lyophilized formulations or aqueoussolutions. Remington: The Science and Practice of Pharmacy 20th Ed.(2000) Lippincott Williams and Wilkins, Ed. K. E. Hoover. Acceptablecarriers, excipients, or stabilizers are nontoxic to recipients at thedosages and concentrations used, and may comprise buffers such asphosphate, citrate, and other organic acids; antioxidants includingascorbic acid and methionine; preservatives (such asoctadecyldimethylbenzyl ammonium chloride; hexamethonium chloride;benzalkonium chloride, benzethonium chloride; phenol, butyl or benzylalcohol; alkyl parabens such as methyl or propyl paraben; catechol;resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecularweight (less than about 10 residues) polypeptides; proteins, such asserum albumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, histidine, arginine, or lysine; monosaccharides,disaccharides, and other carbohydrates including glucose, mannose, ordextrans; chelating agents such as EDTA; sugars such as sucrose,mannitol, trehalose or sorbitol; salt-forming counter-ions such assodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionicsurfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).Pharmaceutically acceptable excipients are further described herein.

In some examples, the pharmaceutical composition described hereincomprises liposomes containing the anti-CεmX antibody, which can beprepared by methods known in the art, such as described in Epstein, etal., Proc. Natl. Acad. Sci. USA 82:3688 (1985); Hwang, et al., Proc.Natl. Acad. Sci. USA 77:4030 (1980); and U.S. Pat. Nos. 4,485,045 and4,544,545. Liposomes with enhanced circulation time are disclosed inU.S. Pat. No. 5,013,556. Particularly useful liposomes can be generatedby the reverse phase evaporation method with a lipid compositioncomprising phosphatidylcholine, cholesterol and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter.

The anti-CεmX antibody may also be entrapped in microcapsules prepared,for example, by coacervation techniques or by interfacialpolymerization, for example, hydroxymethylcellulose orgelatin-microcapsules and poly-(methylmethacylate) microcapsules,respectively, in colloidal drug delivery systems (for example,liposomes, albumin microspheres, microemulsions, nano-particles andnanocapsules) or in macroemulsions. Such techniques are known in theart, see, e.g., Remington, The Science and Practice of Pharmacy 20th Ed.Mack Publishing (2000).

In other examples, the pharmaceutical composition described herein canbe formulated in sustained-release format. Suitable examples ofsustained-release preparations include semipermeable matrices of solidhydrophobic polymers containing the antibody, which matrices are in theform of shaped articles, e.g., films, or microcapsules. Examples ofsustained-release matrices include polyesters, hydrogels (for example,poly(2-hydroxyethyl-methacrylate), or poly(v nylalcohol)), polylactides(U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and 7ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradablelactic acid-glycolic acid copolymers such as the LUPRON DEPOT™(injectable microspheres composed of lactic acid-glycolic acid copolymerand leuprolide acetate), sucrose acetate isobutyrate, andpoly-D-(−)-3-hydroxybutyric acid.

In some embodiments, the pharmaceutical composition comprising theanti-CεmX antibody described herein, e.g., FB825 or a functional variantthereof as also described herein, may be an aqueous formulation, whichmay further comprise a buffer (which may comprise an amino acid such ashistidine or arginine), a salt (e.g., sodium chloride), and/or asurfactant, such as a nonionic surfactant. For example, the aqueousformulation may comprise the antibody at a concentration of about 10-30mg/ml, a buffer comprising an amino acid (e.g., histidine or arginine)at a concentration of about 10-30 mM, a surfactant such as polysorbate80 at a concentration of about 0.01-0.03%, and/or a sodium chloride at aconcentration of about 120-160 mM. Such an aqueous formulation may havea pH of about 5-8. In one particular example, the aqueous formulationmay comprise antibody FB825 at a concentration of about 20 mg/ml,L-histidine at a concentration of about 20 mM, sodium chloride at aconcentration of about 140 mM, polysorbate 80 at a concentration ofabout 0.02%, and a pH of about 6.5.

The term “about” or “approximately” means within an acceptable errorrange for the particular value as determined by one of ordinary skill inthe art, which will depend in part on how the value is measured ordetermined, i.e., the limitations of the measurement system. Forexample, “about” can mean within an acceptable standard deviation, perthe practice in the art. Alternatively, “about” can mean a range of upto ±20%, preferably up to ±10%, more preferably up to ±5%, and morepreferably still up to ±1% of a given value. Alternatively, particularlywith respect to biological systems or processes, the term can meanwithin an order of magnitude, preferably within 2-fold, of a value.Where particular values are described in the application and claims,unless otherwise stated, the term “about” is implicit and in thiscontext means within an acceptable error range for the particular value.

The pharmaceutical compositions to be used for in vivo administrationmust be sterile. This is readily accomplished by, for example,filtration through sterile filtration membranes. Therapeutic antibodycompositions are generally placed into a container having a sterileaccess port, for example, an intravenous solution bag or vial having astopper pierceable by a hypodermic injection needle.

The pharmaceutical compositions described herein can be in unit dosageforms such as tablets, pills, capsules, powders, granules, solutions orsuspensions, or suppositories, for oral, parenteral or rectaladministration, or administration by inhalation or insufflation.

For preparing solid compositions such as tablets, the principal activeingredient can be mixed with a pharmaceutical carrier, e.g.,conventional tableting ingredients such as corn starch, lactose,sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalciumphosphate or gums, and other pharmaceutical diluents, e.g., water, toform a solid preformulation composition containing a homogeneous mixtureof a compound of the present invention, or a non-toxic pharmaceuticallyacceptable salt thereof. When referring to these preformulationcompositions as homogeneous, it is meant that the active ingredient isdispersed evenly throughout the composition so that the composition maybe readily subdivided into equally effective unit dosage forms such astablets, pills and capsules. This solid preformulation composition isthen subdivided into unit dosage forms of the type described abovecontaining from 0.1 to about 500 mg of the active ingredient of thepresent invention. The tablets or pills of the novel composition can becoated or otherwise compounded to provide a dosage form affording theadvantage of prolonged action. For example, the tablet or pill cancomprise an inner dosage and an outer dosage component, the latter beingin the form of an envelope over the former. The two components can beseparated by an enteric layer that serves to resist disintegration inthe stomach and permits the inner component to pass intact into theduodenum or to be delayed in release. A variety of materials can be usedfor such enteric layers or coatings, such materials including a numberof polymeric acids and mixtures of polymeric acids with such materialsas shellac, cetyl alcohol and cellulose acetate.

Suitable surface-active agents include, in particular, non-ionic agents,such as polyoxyethylenesorbitans (e.g., Tween™ 20, 40, 60, 80 or 85) andother sorbitans (e.g., Span™ 20, 40, 60, 80 or 85). Compositions with asurface-active agent will conveniently comprise between 0.05 and 5%surface-active agent, and can be between 0.1 and 2.5%. It will beappreciated that other ingredients may be added, for example mannitol orother pharmaceutically acceptable vehicles, if necessary.

Suitable emulsions may be prepared using commercially available fatemulsions, such as Intralipid™, Liposyn™, Infonutrol™, Lipofundin™ andLipiphysan™. The active ingredient may be either dissolved in apre-mixed emulsion composition or alternatively it may be dissolved inan oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil,corn oil or almond oil) and an emulsion formed upon mixing with aphospholipid (e.g., egg phospholipids, soybean phospholipids or soybeanlecithin) and water. It will be appreciated that other ingredients maybe added, for example glycerol or glucose, to adjust the tonicity of theemulsion. Suitable emulsions will typically contain up to 20% oil, forexample, between 5 and 20%.

The emulsion compositions can be those prepared by mixing an anti-CεmXantibody with Intralipid™ or the components thereof (soybean oil, eggphospholipids, glycerol and water).

Pharmaceutical compositions for inhalation or insufflation includesolutions and suspensions in pharmaceutically acceptable, aqueous ororganic solvents, or mixtures thereof, and powders. The liquid or solidcompositions may contain suitable pharmaceutically acceptable excipientsas set out above. In some embodiments, the compositions are administeredby the oral or nasal respiratory route for local or systemic effect.

Compositions in preferably sterile pharmaceutically acceptable solventsmay be nebulised by use of gases. Nebulised solutions may be breatheddirectly from the nebulising device or the nebulising device may beattached to a face mask, tent or intermittent positive pressurebreathing machine. Solution, suspension or powder compositions may beadministered, preferably orally or nasally, from devices which deliverthe formulation in an appropriate manner.

Use of Anti-CεmX Antibodies for Treating Disorders Associated with IgE

To practice the method disclosed herein, an effective amount of thepharmaceutical composition described above can be administered to asubject (e.g., a human) in need of the treatment via a suitable route,such as intravenous administration, e.g., as a bolus or by continuousinfusion over a period of time, by intramuscular, intraperitoneal,intracerebrospinal, subcutaneous, intra-articular, intrasynovial,intrathecal, oral, inhalation or topical routes. Commercially availablenebulizers for liquid formulations, including jet nebulizers andultrasonic nebulizers are useful for administration. Liquid formulationscan be directly nebulized and lyophilized powder can be nebulized afterreconstitution. Alternatively, anti-CεmX antibodies can be aerosolizedusing a fluorocarbon formulation and a metered dose inhaler, or inhaledas a lyophilized and milled powder.

The subject to be treated by the methods described herein can be amammal, more preferably a human. Mammals include, but are not limitedto, farm animals, sport animals, pets, primates, horses, dogs, cats,mice and rats. A human subject who needs the treatment may be a humanpatient having, at risk for, or suspected of having a disorderassociated with IgE (e.g., allergic asthma, as well as other disordersknown in the art and/or disclosed herein). A subject having anIgE-associated disorder such as allergic asthma can be identified byroutine medical examination, e.g., laboratory tests. A subject suspectedof having the IgE-associated disorder might show one or more symptoms ofthe disorder, e.g., elevated levels of IgE and/or hyper-reactivity to anallergen and/or antigen. A subject at risk for the disorder can be asubject having one or more of the risk factors for that disorder.

Exemplary IgE-associated disorders include, but are not limited to,asthma, allergic rhinitis, hyper IgE syndrome, atopic dermatitis,cold-induced urticaria, chronic urticaria, cholinergic urticaria,chronic rhinosinusitis, systemic mastocytosis, cutaneous mastocytosis,allergic bronchopulmonary aspergillosis, recurrent idiopathicangioedema, and interstitial cystitis, eosinophil-associatedgastrointestinal disorders, a food allergy, or a drug allergy.

“An effective amount” as used herein refers to the amount of each activeagent required to confer therapeutic effect on the subject, either aloneor in combination with one or more other active agents. Effectiveamounts vary, as recognized by those skilled in the art, depending onthe particular condition being treated, the severity of the condition,the individual patient parameters including age, physical condition,size, gender and weight, the duration of the treatment, the nature ofconcurrent therapy (if any), the specific route of administration andlike factors within the knowledge and expertise of the healthpractitioner. These factors are well known to those of ordinary skill inthe art and can be addressed with no more than routine experimentation.It is generally preferred that a maximum dose of the individualcomponents or combinations thereof be used, that is, the highest safedose according to sound medical judgment. It will be understood by thoseof ordinary skill in the art, however, that a patient may insist upon alower dose or tolerable dose for medical reasons, psychological reasonsor for virtually any other reasons.

Empirical considerations, such as the half-life, generally willcontribute to the determination of the dosage. For example, antibodiesthat are compatible with the human immune system, such as humanizedantibodies or fully human antibodies, may be used to prolong half-lifeof the antibody and to prevent the antibody being attacked by the host'simmune system. Frequency of administration may be determined andadjusted over the course of therapy, and is generally, but notnecessarily, based on treatment and/or suppression and/or ameliorationand/or delay of a disorder associated with IgE. Alternatively, sustainedcontinuous release formulations of an anti-CεmX antibody may beappropriate. Various formulations and devices for achieving sustainedrelease are known in the art.

In one example, dosages for an anti-CεmX antibody as described hereinmay be determined empirically in individuals who have been given one ormore administration(s) of an anti-CεmX antibody. Individuals are givenincremental dosages of the anti-CεmX antibody. To assess efficacy of theanti-CεmX antibody, an indicator of a disorder associated with IgE (suchas levels of IgE) can be followed.

For the purpose of the present disclosure, the appropriate dosage of ananti-CεmX antibody will depend on the specific anti-CεmX antibody(s) (orcompositions thereof) employed, the type and severity of disorderassociated with IgE, whether the antibody is administered for preventiveor therapeutic purposes, previous therapy, the patient's clinicalhistory and response to the antibody, and the discretion of theattending physician. Typically the clinician will administer ananti-CεmX antibody, such as FB825, until a dosage is reached thatachieves the desired result. Administration of an anti-CεmX antibody canbe continuous or intermittent, depending, for example, upon therecipient's physiological condition, whether the purpose of theadministration is therapeutic or prophylactic, and other factors knownto skilled practitioners.

In some embodiments, the anti-CεmX antibody (e.g., FB825) describedherein is administered to a subject in need of the treatment at anamount sufficient to reduce the level of the total IgE level by at least20% (e.g., 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater).

As used herein, the term “treating” refers to the application oradministration of a composition including one or more active agents to asubject, who has a disease associated with IgE, a symptom of a diseaseassociated with IgE, or a predisposition toward the disease, with thepurpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate,improve, or affect the disorder, the symptom of the disease, or thepredisposition toward the disease.

Alleviating a disease associated with IgE includes delaying thedevelopment or progression of the disease, or reducing disease severity.Alleviating the disease does not necessarily require curative results.As used therein, “delaying” the development of a disease (such as adisease associated with IgE) means to defer, hinder, slow, retard,stabilize, and/or postpone progression of the disease. This delay can beof varying lengths of time, depending on the history of the diseaseand/or individuals being treated. A method that “delays” or alleviatesthe development of a disease, or delays the onset of the disease, is amethod that reduces probability of developing one or more symptoms ofthe disease in a given time frame and/or reduces extent of the symptomsin a given time frame, when compared to not using the method. Suchcomparisons are typically based on clinical studies, using a number ofsubjects sufficient to give a statistically significant result.

“Development” or “progression” of a disease means initial manifestationsand/or ensuing progression of the disease. Development of the diseasecan be detectable and assessed using standard clinical techniques aswell known in the art. However, development also refers to progressionthat may be undetectable. For purpose of this disclosure, development orprogression refers to the biological course of the symptoms.“Development” includes occurrence, recurrence, and onset. As used herein“onset” or “occurrence” of a disease associated with IgE includesinitial onset and/or recurrence.

To perform the methods as described herein, any of the anti-Cεmxantibodies such as FB825 may be given to a subject in need of thetreatment (e.g., a human patient) by a single dose or by multiple dosesvia a suitable route, for example, intravenous infusion or subcutaneousinjection. The dosage of the anti-Cεmx antibody for each administrationmay range from about 0.5 mg/kg to about 25 mg/kg (e.g., about 1 mg/kg toabout 20 mg/kg, about 5 mg/kg to about 15 mg/kg, or about 10 mg/kg toabout 20 mg/kg), depending upon various factors, including thosedescribed herein. For repeated administrations over several days orlonger, depending on the condition, the treatment is sustained until adesired suppression of symptoms occurs or until sufficient therapeuticlevels are achieved to alleviate a disorder associated with IgE, or asymptom thereof.

The administration of an anti-CεmX antibody (e.g., FB825) may beessentially continuous over a preselected period of time or may be in aseries of spaced dose, e.g., either before, during, or after developinga disorder associated with IgE. An exemplary dosing regimen comprisesadministering to a subject in need of the treatment a first dose of ananti-Cεmx antibody (e.g., at 3 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20mg/kg, or 25 mg/kg), followed by a second dose of the antibody at least3 months after the first dose (e.g., 4 months, 5 months, or 6 months).The dosage of the second administration may be higher, the same, orlower than the first administration. Other dosage regimens may be usefuldepending upon the pattern of pharmacokinetic decay that a practitionerwishes to achieve.

In some embodiments, a subject in need of the treatment can be given afirst dose of the antibody at a suitable amount (e.g., at 3 mg/kg, 5mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, or 25 mg/kg). The subject is thenmonitored periodically for symptoms indicative of an IgE-associateddisorder, for example, allergic reactions and/or an elevated level oftotal IgE. A second dose of the antibody may be given to the subjectwhen such a symptom is observed.

Also within the scope of the present disclosure are preventivetreatments of an IgE-associated disorder with any of the anti-Cεmxantibodies to reduce the risk for occurrence of such a disorder.Subjects suitable for such a preventive treatment may be human patientshaving history of an IgE-associated disorder and/or family history of anIgE-associated disorder.

Conventional methods, known to those of ordinary skill in the art ofmedicine, can be used to administer the pharmaceutical composition tothe subject, depending upon the type of disease to be treated or thesite of the disease. This composition can also be administered via otherconventional routes, e.g., administered orally, parenterally, byinhalation spray, topically, rectally, nasally, buccally, vaginally orvia an implanted reservoir. The term “parenteral” as used hereinincludes subcutaneous, intracutaneous, intravenous, intramuscular,intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal,intralesional, and intracranial injection or infusion techniques. Inaddition, it can be administered to the subject via injectable depotroutes of administration such as using 1-, 3-, or 6-month depotinjectable or biodegradable materials and methods.

Injectable compositions may contain various carriers such as vegetableoils, dimethylactamide, dimethyformamide, ethyl lactate, ethylcarbonate, isopropyl myristate, ethanol, and polyols (glycerol,propylene glycol, liquid polyethylene glycol, and the like). Forintravenous injection, water soluble antibodies can be administered bythe drip method, whereby a pharmaceutical formulation containing theantibody and a physiologically acceptable excipients is infused.Physiologically acceptable excipients may include, for example, 5%dextrose, 0.9% saline, Ringer's solution or other suitable excipients.Intramuscular preparations, e.g., a sterile formulation of a suitablesoluble salt form of the antibody, can be dissolved and administered ina pharmaceutical excipient such as Water-for-Injection, 0.9% saline, or5% glucose solution.

In one embodiment, an anti-CεmX antibody is administered viasite-specific or targeted local delivery techniques. Examples ofsite-specific or targeted local delivery techniques include variousimplantable depot sources of the anti-CεmX antibody or local deliverycatheters, such as infusion catheters, an indwelling catheter, or aneedle catheter, synthetic grafts, adventitial wraps, shunts and stentsor other implantable devices, site specific carriers, direct injection,or direct application. See, e.g., PCT Publication No. WO 00/53211 andU.S. Pat. No. 5,981,568. Targeted delivery of therapeutic compositionscontaining an antisense polynucleotide, expression vector, or subgenomicpolynucleotides can also be used. Receptor-mediated DNA deliverytechniques are described in, for example, Findeis et al., TrendsBiotechnol. (1993) 11:202; Chiou et al., Gene Therapeutics: Methods AndApplications Of Direct Gene Transfer (J. A. Wolff, ed.) (1994); Wu etal., J. Biol. Chem. (1988) 263:621; Wu et al., J. Biol. Chem. (1994)269:542; Zenke et al., Proc. Natl. Acad. Sci. USA (1990) 87:3655; Wu etal., J. Biol. Chem. (1991) 266:338. Therapeutic compositions containinga polynucleotide are administered in a range of about 100 ng to about200 mg of DNA for local administration in a gene therapy protocol. Insome embodiments, concentration ranges of about 500 ng to about 50 mg,about 1 μg to about 2 mg, about 5 μg to about 500 μg, and about 20 μg toabout 100 μg of DNA or more can also be used during a gene therapyprotocol.

The therapeutic polynucleotides and polypeptides described herein can bedelivered using gene delivery vehicles. The gene delivery vehicle can beof viral or non-viral origin (see generally, Jolly, Cancer Gene Therapy(1994) 1:51; Kimura, Human Gene Therapy (1994) 5:845; Connelly, HumanGene Therapy (1995) 1:185; and Kaplitt, Nature Genetics (1994) 6:148).Expression of such coding sequences can be induced using endogenousmammalian or heterologous promoters and/or enhancers. Expression of thecoding sequence can be either constitutive or regulated.

Viral-based vectors for delivery of a desired polynucleotide andexpression in a desired cell are well known in the art. Exemplaryviral-based vehicles include, but are not limited to, recombinantretroviruses (see, e.g., PCT Publication Nos. WO 90/07936; WO 94/03622;WO 93/25698; WO 93/25234; WO 93/11230; WO 93/10218; WO 91/02805; U.S.Pat. Nos. 5,219,740 and 4,777,127; GB Patent No. 2,200,651; and EPPatent No. 0 345 242), alphavirus-based vectors (e.g., Sindbis virusvectors, Semliki forest virus (ATCC VR-67; ATCC VR-1247), Ross Rivervirus (ATCC VR-373; ATCC VR-1246) and Venezuelan equine encephalitisvirus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-532)), andadeno-associated virus (AAV) vectors (see, e.g., PCT Publication Nos. WO94/12649, WO 93/03769; WO 93/19191; WO 94/28938; WO 95/11984 and WO95/00655). Administration of DNA linked to killed adenovirus asdescribed in Curiel, Hum. Gene Ther. (1992) 3:147 can also be employed.Non-viral delivery vehicles and methods can also be employed, including,but not limited to, polycationic condensed DNA linked or unlinked tokilled adenovirus alone (see, e.g., Curiel, Hum. Gene Ther. (1992)3:147); ligand-linked DNA (see, e.g., Wu, J. Biol. Chem. (1989)264:16985); eukaryotic cell delivery vehicles cells (see, e.g., U.S.Pat. No. 5,814,482; PCT Publication Nos. WO 95/07994; WO 96/17072; WO95/30763; and WO 97/42338) and nucleic charge neutralization or fusionwith cell membranes. Naked DNA can also be employed. Exemplary naked DNAintroduction methods are described in PCT Publication No. WO 90/11092and U.S. Pat. No. 5,580,859. Liposomes that can act as gene deliveryvehicles are described in U.S. Pat. No. 5,422,120; PCT Publication Nos.WO 95/13796; WO 94/23697; WO 91/14445; and EP Patent No. 0524968.Additional approaches are described in Philip, Mol. Cell. Biol. (1994)14:2411, and in Woffendin, Proc. Natl. Acad. Sci. (1994) 91:1581.

It is also apparent that an expression vector can be used to directexpression of any of the protein-based anti-CεmX antibodies describedherein (e.g., FB825). For example, other anti-CεmX antibody fragmentsthat are capable of binding CεmX and/or an IgE biological activity areknown in the art.

The particular dosage regimen, i.e., dose, timing and repetition, usedin the method described herein will depend on the particular subject andthat subject's medical history. Any of the anti-Cmex antibodiesdescribed herein may be used in conjunction with other agents (e.g.,other agents for treating IgE-associated disorders) that serve toenhance and/or complement the effectiveness of the agents.

In some embodiments, an anti-CεmX antibody as described herein, forexample, FB825, is used for treating atopic dermatitis as follows.Atopic dermatitis, also known as eczema, is a chronical skin conditioncharacterized by redness and/or itchy. It is common in children but canoccur at any age. A patient who needs the treatment can be identified byroutine medical practice as having one or more symptoms of atopicdermatitis, including dry skin, itching, red to brownish-gray patches,small, raised bumps, which may leak fluid and crust over when scratched,thickened, cracked, scaly skin, and/or raw, sensitive, swollen skin fromscratching. In some instances, the total IgE level and the level ofallergen-specific IgE of a candidate subject can be examined via routinepractice. If the IgE level of the candidate subject (e.g., the totalIgE, the allergen-specific IgE, or both) is higher than a normal level(representing the average IgE level in subjects of the same species,e.g., humans, who are free of atopic dermatitis or other allergicdisorders associated with IgE).

A human patient who needs the treatment may be given a first a dose ofthe antibody, which may range from 3 mg/kg to 8 mg/kg, via aconventional route as described herein. In some instances, the firstdose is 5 mg/kg. After the first dose, the total IgE level of thepatient can be monitored. If the reduction of the IgE level 3-4 weeksafter the first dose is less than 50%, a second dose of the antibody maybe given to the patient 3-4 weeks after the first dose. The second dosemay be identical to the first dose, or lower than the first dose. Insome instances, both the first dose and the second dose are 5 mg/kg andare administered via IV infusion in a 1-2 hour period. Other biomarkersindicating efficacy and/or safety could also be monitored during thecourse of the treatment. Such biomarkers include, but are not limitedto, thymus and activation regulated chemokine (TARC), Eotaxin-3, thymicstromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13,IL-16, IL-31, M-CSF, or a combination thereof.

The human patient subject to the above-noted treatment may have chronicatopic dermatitis for at least 3 years as diagnosed by routine medicalpractice, for example, defined by the Eishenfield revised criteria ofHannifin and Rajka and supported by positive allergen-specific IgE. Thepatient may have one or more of the following features: (i) eczema areaand severity index (EASI) score greater than 14, (ii) Investigator'sGlobal Assessment (IGA) score greater than 3 (5-point scale), (iii)greater than 10% body surface area (BSA), (iv) history of inadequateresponse to a stable regimen of topical corticosteroids or calcineurininhibitors for at least one month or at least three months before thetreatment. Further, the human patient may be given stable doses ofemollient twice daily for at least 7 days before the treatment.

In some instances, the anti-CεmX antibody as described herein (e.g.,FB825) may be co-used with moisturizers (e.g., at stable doses such asat least twice daily) and/or topical corticosteroid (TCS). A mediumpotency TCS may be applied to areas with active lesions and may switchto low potency TCS after the lesions are under control. If lesionsreoccur, treatment with a medium potency TCS may resume with a step-downapproach. If lesions are persisting or worsening after daily treatmentwith a medium potency TCS, a high or super-high potency TCS may be used,unless it is deemed unsafe. A low potency TCS may be used on areas ofthin skin (e.g., face, neck, intertriginous, genital areas, or areas ofskin atrophy) or on areas where continued use of medium potency TCS isconsidered unsafe.

TCS having low, medium, and high or super-high potency is well known inthe art. Exemplary medium potency TCS includes 0.05% fluticasonepropionate cream, 0.1% mometasone furoate cream, or 0.06% betamethasonevalerate cream. Exemplary low potency TCS includes 1% hydrocortisoneointment. Exemplary high potency TCS can be 0.05% fluocinonide cream or0.25% desomimetasone ointment. Exemplary super-high potency TCS can be0.05% clobetasol propionate ointment.

In some instances, the patient subject to the treatment described hereinis free of one or more of the following therapy: (i) topical tacrolimusand pimecrolimus, (ii) systemic treatment of corticosteroids, (iii)leukotriene inhibitors, (iv) allergen immunotherapy, (v) systemictreatment of immunosuppressors or immunomodulators (e.g., cyclosporine,mycophenolate-mofetil, azathioprine, methotrexate, or biologics), (vi)live (e.g., attenuated) vaccines, and/or (vii) traditional Chinesemedicine. The patient may also be free of any surgical procedures and/orUV procedures.

Any of the methods described herein may further comprise assessingoccurrence of decreased hemoglobin, upper respiratory tract infection,urinary tract infection, or a combination thereof in the subject afterthe first dose. If one or more occurrences are observed, the amount ofthe anti-Cεmx antibody (e.g., FB825) of the second dose may be reduced.Alternatively, the treatment may be stopped.

Kits for Use in Treating IgE-Associated Disorders

The present disclosure also provides kits for use in treatingIgE-associated disorders. Such kits can include one or more containerscomprising an anti-Cεmx antibody as described herein (such as FB825).

In some embodiments, the kit can comprise instructions for use inaccordance with any of the methods described herein. The includedinstructions can comprise a description of administration of theanti-Cεmx antibody to treat, delay the onset, or alleviate anIgE-associated disorder according to any of the methods describedherein. The kit may further comprise a description of selecting anindividual suitable for treatment based on identifying whether thatindividual has, is suspected of having, or is at risk for the disorder.In still other embodiments, the instructions comprise a description ofadministering anti-Cεmx antibody to a subject in need of the treatmentto reduce the risk for developing the IgE-associated disorder.

The instructions relating to the use of an anti-Cεmx antibody generallyinclude information as to dosage, dosing schedule, and route ofadministration for the intended treatment. The containers may be unitdoses, bulk packages (e.g., multi-dose packages) or sub-unit doses.Instructions supplied in the kits of the invention are typically writteninstructions on a label or package insert (e.g., a paper sheet includedin the kit), but machine-readable instructions (e.g., instructionscarried on a magnetic or optical storage disk) are also acceptable.

The label or package insert indicates that the composition is used fortreating, delaying the onset and/or alleviating an IgE-associateddisorder. Instructions may be provided for practicing any of the methodsdescribed herein.

The kits of this disclosure are in suitable packaging. Suitablepackaging includes, but is not limited to, vials, bottles, jars,flexible packaging (e.g., sealed Mylar or plastic bags), and the like.Also contemplated are packages for use in combination with a specificdevice, such as an inhaler, nasal administration device (e.g., anatomizer) or an infusion device such as a minipump. A kit may have asterile access port (for example the container may be an intravenoussolution bag or a vial having a stopper pierceable by a hypodermicinjection needle). The container may also have a sterile access port(for example the container may be an intravenous solution bag or a vialhaving a stopper pierceable by a hypodermic injection needle). At leastone active agent in the composition is an anti-Cεmx antibody, such asFB825.

Kits may optionally provide additional components such as buffers andinterpretive information. Normally, the kit comprises a container and alabel or package insert(s) on or associated with the container. In someembodiments, the present disclosure provides articles of manufacturecomprising contents of the kits described above.

General Techniques

The practice of the present invention will employ, unless otherwiseindicated, conventional techniques of molecular biology (includingrecombinant techniques), microbiology, cell biology, biochemistry andimmunology, which are within the skill of the art. Such techniques areexplained fully in the literature, such as, Molecular Cloning: ALaboratory Manual, second edition (Sambrook, et al., 1989) Cold SpringHarbor Press; Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Methodsin Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook(J. E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R. I.Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J. P.Mather and P. E. Roberts, 1998) Plenum Press; Cell and Tissue Culture:Laboratory Procedures (A. Doyle, J. B. Griffiths, and D. G. Newell,eds., 1993-8) J. Wiley and Sons; Methods in Enzymology (Academic Press,Inc.); Handbook of Experimental Immunology (D. M. Weir and C. C.Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J. M.Miller and M. P. Calos, eds., 1987); Current Protocols in MolecularBiology (F. M. Ausubel, et al., eds., 1987); PCR: The Polymerase ChainReaction, (Mullis, et al., eds., 1994); Current Protocols in Immunology(J. E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology(Wiley and Sons, 1999); Immunobiology (C. A. Janeway and P. Travers,1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D.Catty., ed., IRL Press, 1988-1989); Monoclonal antibodies: a practicalapproach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000);Using antibodies: a laboratory manual (E. Harlow and D. Lane (ColdSpring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995).

Without further elaboration, it is believed that one skilled in the artcan, based on the above description, utilize the present invention toits fullest extent. The following specific embodiments are, therefore,to be construed as merely illustrative, and not limitative of theremainder of the disclosure in any way whatsoever. All publicationscited herein are incorporated by reference for the purposes or subjectmatter referenced herein.

Example 1: Toxicity Studies of FB825 in Cynomolgus Monkey Materials andMethods Laboratory Tests

Blood and urine samples for hematology, coagulation, serum chemistry(including liver function tests), thyroid function tests, and urinalysiswere collected and analyzed following routine clinical laboratory tests.

Abnormal clinical laboratory values were flagged as either high or low(or normal or abnormal) based on the reference ranges for eachlaboratory parameter. Clinical significance was defined as any variationin results that had medical relevance and may have resulted in analteration in medical care (e.g., active observation, diagnosticmeasures, or therapeutic measures). If a clinically significant changefrom screening was noted, the clinically significant value and reasonfor clinical significance were documented. The cynomolgus monkey beingtreated was continued to be monitored with additional assessments untilthe values reached the reference range or the values at screening oruntil follow-up was no longer medically necessary.

Pharmacodynamic Assessments

Blood samples for the determination of total IgE and antidrug antibodies(ADA) were collected using 3.5-mL blood collection tubes (BD Vacutainer®SST™ Serum Separation Tubes), provided by Vince and Associates ClinicalResearch. One sample was collected to target a minimum blood volume perblood collection tube size. A minimum of 1.0 mL of serum was collectedat each time point. After obtaining the blood sample, the collectiontube was inverted 5 times and the blood was allowed to clot for 30minutes at ambient temperature (19° C.-24° C.). The sample wascentrifuged at approximately 2200 rpm for 10 minutes at room temperaturein a swinging bucket centrifuge. Duplicate serum aliquots ofapproximately equal volume (minimum of 500 μL per aliquot) weretransferred, using the standard laboratory technique, into 2appropriately labeled storage tubes (2-mL polypropylene cryovials)provided by Vince and Associates Clinical Research.

Immunogenicity samples for the measurement of ADA were analyzed using avalidated ELISA.

Serum Aspartate Aminotransferase and Alanine Amino Transferase

The liver function in FB825 treated cynomolgus monkeys was monitored bymeasuring the levels of aspartate aminotransferase (AST) and alanineamino transferase (ALT). The activities of these two enzymes wereexpressed as unit per liter (U/L).

Results Single-Dose Studies of FB825 in Cynomolgus Monkeys

Single-dose toxicity of FB825 was assessed in non-GLP single-dosetoxicity studies carried out in cynomolgus monkeys. In the first study,treatment-related effects were determined in cynomolgus monkeysadministered a single 10-minute IV infusion of FB825. In the secondstudy, treatment-related effects were determined in cynomolgus monkeysadministered a single subcutaneous injection of FB825.

In an IV injection dose-range-finding study, a single 10-minute IVinfusion of FB825 was well tolerated in male and female cynomolgusmonkeys at 30, 100, and 300 mg/kg. There were no treatment-relatedeffects on clinical parameters, food consumption, body weight, ormortality. FB825-related effects were limited to minimal increases inALT and AST that had recovered by Day 57 and minimal increases ininterleukin-6 and interleukin-10 at 6 hours and/or 1 hour after dosingin animals administered at least 100 mg/kg FB825. Based on theseresults, the no observed adverse effect level (NOAEL) was considered tobe 300 mg/kg.

In another single-dose toxicology study, administration of FB825 via asingle subcutaneous injection was well tolerated in cynomolgus monkeysat 300 mg/kg. No FB825-related clinical signs or effects on body weightsor clinical pathology parameters (hematology, coagulation, and clinicalchemistry) were observed.

Repeat-Dose Studies of FB825 in Cynomolgus Monkeys

Repeat-dose toxicity of FB825 was assessed in cynomolgus monkeys.Treatment-related effects were determined in cynomolgus monkeysadministered a single 10-minute IV infusion of FB825 once weekly for atotal of 4 doses.

Administration of FB825 by 10 minute IV infusion once weekly for a totalof 4 doses was well tolerated in cynomolgus monkeys at 30, 100, and 300mg/kg. The following parameters and end points were evaluated in thisstudy: clinical signs, body weight, food consumption, ophthalmology,electrocardiology, clinical pathology parameters (hematology,coagulation, and clinical chemistry), bioanalysis and toxicokineticevaluations, antitherapeutic antibody analysis, flow cytometry, IgEanalysis, thyroid hormone levels, gross necropsy findings, organweights, and histopathologic examinations.

No FB825-related clinical signs or effects observed on body weight, foodconsumption, the ophthalmic and electrocardiographic examinations,coagulation parameters, and thyroid hormone levels were observed ineither sex at doses up to 300 mg/kg. Additionally, there were noFB825-related notable gross findings at doses up to and including 300mg/kg.

FB825-related effects were limited to reversible marked increases in ALTat FB825 doses of higher than or equal to 100 mg/kg, and reversible mildincreases in AST, partially reversible minimal decreases in albuminlevels and the corresponding albumin:globulin ratios, and reversibleminimal increases in monocyte counts at an FB825 dose of 300 mg/kg. Themagnitude of the increases in ALT and AST levels at 300 mg/kg wereconsidered adverse. Target organ effects were observed at levels of ≥30mg/kg and consisted of nonadverse follicular colloid depletion of thethyroid and lower thyroid weight. However, the lower thyroid weightswere within the range observed in historical control monkeys. Becausevariation in colloid staining pattern, variation in thyroid folliclesize, and vacuolar degeneration have been reported as spontaneousfindings in cynomolgus monkeys (Ishida 2000; Hatakeyama 2011) andbecause there were no test article-related effects on thyroxine andthyroid-stimulating hormone levels, the findings in the thyroid wereconsidered nonadverse under the conditions of this study.

Based on the increases in ALT and AST levels at 300 mg/kg, the NOAEL wasconsidered to be 100 mg/kg (C_(max) of 5330 μg/mL and AUC_((0-168 h)) of520 mg·hr/mL for males and C_(max) of 5220 μg/mL and AUC_((0-168 h)) of487 mg·hr/mL for females).

Example 2: Human Clinical Studies

The primary objective of this study was to evaluate the safety andtolerability of single ascending IV doses of FB825 in normal, healthysubjects. The secondary objectives of this study include determinationthe PK profile of single ascending IV doses of FB825, to explore theeffects on total IgE after single ascending IV doses of FB825, and toexplore the occurrence of anti-FB825 antibodies after single ascendingIV doses of FB825.

Study Design

This was a Phase 1, first-in-human (FIH), randomized, double-blind,placebo-controlled study to evaluate the safety, tolerability,pharmacokinetics, and immunogenicity of single ascending IV doses ofFB825. An overview of the schedule of events performed during the FB825human clinical study is described in Table 1. Subjects who met thecriteria for study entry were assigned to the current dose cohort andrandomly assigned to receive FB825 or placebo (vehicle). All doses ofFB825 were administered as a 1-hour IV infusion.

The safety data for each dose cohort were reviewed before progression tothe next-highest-dose cohort. Dosing for the next-highest-dose cohortwas permitted only after confirmation of the safety to do so. A doselevel could have been adjusted, or repeated, as necessary. Blinded PKdata was reviewed at the same time as the blinded safety data.

The study included 6 cohorts and a total of approximately 54 normal,healthy subjects (7 subjects each [4 active: 3 placebo] in Cohorts A andB and 10 subjects each [7 active: 3 placebo] in Cohorts C, D, E, and F).The starting FB825 dose was 0.003 mg/kg IV with planned increases insubsequent cohorts to 0.03, 0.3, 1.5, 5, and 10 mg/kg IV. The studyconsisted of a screening period (Days −28 to −2), check-in (Day −1), atreatment/follow-up period (Days 1 to 140), and an end-of-study visit(Day 140).

Subjects checked into the clinic on Day −1, and check-in procedures wereperformed. When procedures overlapped and were scheduled to occur at thesame time point, the order of procedures was vital sign measurements,electrocardiograms, and then pharmacokinetic blood collection.

Subjects received active or placebo study drug on Day 1. The first 2subjects from the 2 lowest-dose cohorts (Cohorts A and B) receivedeither placebo or FB825 (i.e., both placebo and active study drug wererequired to be represented). The remaining subjects in these cohortswere dosed 48 hours after the first 2 subjects were dosed. All subjectsin the 4 highest-dose cohorts were dosed at the same time. There was aminimum of 14 days between the dosing of the last subject in a cohortbefore a decision was made to proceed with dosing the next cohort.Safety data for each dose cohort were reviewed in a blinded fashionbefore progression to the next-highest-dose cohort. Dosing in thenext-highest-dose cohort was permitted only after confirmation that itwas safe to do so. Blinded PK data was reviewed at the same time as theblinded safety data.

Blood samples for PK assessments were collected on Day 1 at 30 minutes(±5 minutes) before the start of infusion; at 30 minutes (±2 minutes)after the start of infusion; at 1, 1.25, and 2 hours (±2 minutes) afterthe start of infusion; at 4 and 8 hours (±5 minutes) after the start ofinfusion; and at 24 and 48 hours (±10 minutes) after the start ofinfusion. In addition, single blood samples were collected on Days 5,14, 29, 85, and 140 after infusion. Blood samples for immunogenicityassessments were collected on Day 1 (30 minutes [±5 minutes] before thestart of infusion) and on Days 5, 14, 29, 85, and 140.

Subjects were confined within the clinic from Day −1 until discharge onDay 3 (48 hours after dosing) and returned to the clinic on Days 5, 14,29, 85, and 140 for outpatient visits. The duration of the study,excluding screening, was approximately 140 days.

Patient Population

Healthy male and female subjects, 18 to 55 years of age, inclusive, hada body weight higher than or equal to 50 kg and a body mass index of18.0 to 30.0 kg/m2, inclusive, and provided written informed consent. Atotal of 54 subjects were enrolled, 41 subjects (75.9%) completed thestudy, and 13 subjects (24.1%) were discontinued. Seven subjects (13.0%)discontinued due to subject choice and included 1 subject each in the0.3, 1.5, 5, and 10 mg/kg FB825 treatment groups and 3 subjects in theplacebo treatment group. Six subjects (11.1%) were lost to follow-up andincluded 1 subject each in the 0.003, 0.3, and 1.5 mg/kg FB825 andplacebo treatment groups and 2 subjects in the 10 mg/kg FB825 treatmentgroup.

TABLE 1 Schedule of Events. Procedure Confinement 2 3 Outpatient VisitsCheck- (24 h (48 h 14 29 85 140 Screening in after after (+/−1 (+/−1(+/−1 (EOS) Study Day(s) −28 to −2 −1 1 dosing) dosing) 5 day) day) day)(+/−1 day) Informed consent X Inclusion/Exclusion X X criteria Medicalhistory X X Physical examination X X X X X X X X Vital sign measurementsX X X X X X X X X X 12-Lead ECG X X X X X X X X Cardiac telemetry XSerology X Clinical laboratory X X X X X X X X X X testing Thyroidfunction testing X X X X X X Urine, drug, cotinine, X X and alcoholscreen Serum pregnancy test X X X X (female subjects) Serum FSH XAdmission to clinic X Randomization X Study drug X administrationPharmacokinetic blood X X X X X X X X sampling Total IgE and ADA X X X XX X sampling Adverse events X X X X X X X X assessment Prior andconcomitant X X X X X X X X X X medications Discharge from clinic XStudy discharge X Abbreviations: ADA, antidrug antibodies; ECG,electrocardiogram; EOS, endofstudy; FSH, follicle-stimulating hormone;Ig, immunoglobulin.

FB825 Administration

On Day 1, subjects received a single IV infusion of FB825 (0.003, 0.03,0.3, 1.5, 5, or 10 mg/kg) or placebo over approximately 1 hour.

All doses were administered in the morning of the scheduled dosing day(Day 1) as a single, approximately 1-hour IV infusion after a fast ofapproximately 2 hours. A light breakfast was permitted 2 hours or morebefore dosing. Water was permitted at any time except for 1 hour beforeand 1 hour after dosing. The FB825 solutions were diluted beforeadministration.

A single, approximately 1-hour intravenous infusion of FB825 or placebowas administered on Day 1 at 0 hours. Progress to the next dose levelwas dependent on Safety Review Team approval. The first 2 subjects fromthe 2 lowest-dose cohorts (Cohorts A and B) received either placebo orFB825 (i.e., both placebo and active were required to be represented).The remaining subjects in these cohorts were dosed 48 hours after thefirst 2 subjects had been dosed. All subjects in the 4 highest-dosecohorts were dosed at the same time.

Pharmacokinetic (PK) Assessments

Blood samples for PK analysis of FB825 were collected from all subjectsat the following time points: on Day 1 at 30 minutes (±5 minutes) beforethe start of infusion; at 30 minutes (±2 minutes) after the start ofinfusion; at 1, 1.25, and 2 hours (±2 minutes for 30 minutes to 2 hourspostdose) after the start of infusion; at 4 and 8 hours (±5 minutes for4 to 8 hours postdose) after the start of infusion; and at 24 and 48hours (±10 minutes for 24 to 48 hours postdose) after the start ofinfusion. In addition, single blood samples were collected on Days 5, 14(±1 day), 29 (±2 days), 85 (±3 days), and 140 (±5 days) after infusion.

The following single-dose PK parameters were calculated for FB825 fromthe serum concentration data for each subject using noncompartmentalmethods:

-   -   AUC_(0-t) Area under the serum concentration-time curve (AUC)        from time 0 to the last quantifiable concentration, calculated        using the linear trapezoidal rule    -   AUC_(0-inf) AUC from time 0 extrapolated to infinity, calculated        using the following formula:

AUC_(0-inf)=AUC_(0-t) +C _(t) /K _(el),

-   -   -   where C_(t) was the last measurable serum concentration, and            K_(el) was the terminal elimination rate constant. If the            extrapolated area (C_(t)/K_(el)) was greater than 20% of            AUC_(0-inf), then AUC_(0-inf) and its associated parameters            (CL and V_(d)) were set to missing.

    -   % AUC_(ex) Percentage of the area extrapolated for calculation        of AUC_(0-inf)

    -   C_(max) Maximum observed serum concentration

    -   T_(max) Time of maximum observed serum concentration

    -   K_(el) Terminal elimination rate constant, where K_(el) was the        magnitude of the slope of the linear regression of the log        concentration versus time profile during the terminal phase.        K_(el) was only retained if R²≥0.80 and 3 points in the terminal        phase did not include C_(max).

    -   t_(1/2) Terminal half-life (whenever possible), calculated as        (ln)2/K_(el)

    -   MRT Mean residence time, calculated as AUMC/AUC

    -   CL Apparent clearance, calculated as Dose/AUC_(0-inf)

    -   V_(d) Apparent volume of distribution, calculated as follows:

Dose/(AUC_(0-inf) ×K _(el))

Pharmacokinetic Blood Samples

Blood samples to analyze FB825 were collected using 3.5 mL bloodcollection tubes (BD Vacutainer® SST™ serum separation tubes), providedby Vince and Associates Clinical Research. One sample was collected totarget a minimum blood volume per blood collection tube size. A minimumof 1.0 mL of serum was collected at each time point. After obtaining theblood sample, the collection tube was inverted 5 times, and the bloodwas allowed to clot for 30 minutes at ambient temperature (19° C.-24°C.). The sample was centrifuged at approximately 2200 rpm for 10 minutesat room temperature in a swinging bucket centrifuge. Duplicate serumaliquots of approximately equal volume (minimum of 500 μL per aliquot),using standard laboratory technique, were transferred into 2appropriately labeled storage tubes (2-mL polypropylene cryovials)provided by Vince and Associates Clinical Research. A label was securedto each storage tube and contained the following information:

-   -   Sample Type: Human serum    -   Assay Type: PK    -   Protocol: FB825CLCT01    -   Subject Number: Randomization number    -   Time point: Refer to protocol    -   Serum Aliquot: 1 or 2

Within 90 minutes of collection, both aliquot samples were storedupright at −70° C.±10° C. Pharmacokineticserum determinations of FB825were performed using a validated ELISA.

Pharmacodynamic Assessments

Blood samples for the determination of total IgE and antidrug antibodies(ADA) were collected on Day 1 at 30 minutes (±5 minutes) before thestart of infusion and on Days 5, 14 (±1 day), 29 (±2 days), 85 (±3days), and 140 (±5 days) after the infusion, or at early termination.

Blood samples for analysis of the ADA levels were collected using 3.5-mLblood collection tubes (BD Vacutainer® SST™ Serum Separation Tubes),provided by Vince and Associates Clinical Research. One sample wascollected to target a minimum blood volume per blood collection tubesize. A minimum of 1.0 mL of serum was collected at each time point.After obtaining the blood sample, the collection tube was inverted 5times and the blood was allowed to clot for 30 minutes at ambienttemperature (19° C.-24° C.). The sample was centrifuged at approximately2200 rpm for 10 minutes at room temperature in a swinging bucketcentrifuge. Duplicate serum aliquots of approximately equal volume(minimum of 500 μL per aliquot) were transferred, using the standardlaboratory technique, into 2 appropriately labeled storage tubes (2-mLpolypropylene cryovials). Immunogenicity samples for the measurement ofADA were analyzed using a validated ELISA.

Safety Assessments

Safety and tolerability were assessed by monitoring and recording AEs,clinical laboratory test results (hematology, coagulation, serumchemistry including liver function tests, thyroid function tests, andurinalysis), vital sign measurements, 12-lead ECG results, cardiactelemetry data, and physical examination findings.

Clinical Laboratory Tests

Blood and urine samples for hematology, coagulation, serum chemistry(including liver function tests), thyroid function tests, urinalysis,and drug screen tests were collected under fasting conditions (fastedfor approximately 2 or more hours) at the time points indicated in theschedule of events (Table 1). Clinical laboratory tests (hematology,coagulation, serum chemistry [including liver function tests], andurinalysis) were performed at screening; check-in; on Day 1 before thestart of infusion and at 1 (end of infusion), 8, and 24 hours (±15minutes) after the start of the infusion; and at a single time point onDays 3 (±15 minutes for up to 48 hours postdose), 5, 14, 29, 85, and140.

The samples were used for clinical laboratory tests includinghematology, coagulation, serum chemistry, thyroid function, andurinalysis.

A serum pregnancy test (β-human chorionic gonadotropin) was performed onall female subjects at screening, check-in, on Day 3 (48 hours afterdosing), and at the end-of-study visit (Day 140). Female subjects whowere postmenopausal had a serum FSH test at screening.

Hepatitis B surface antigen, hepatitis C virus antibody, and humanimmunodeficiency virus (types 1 or 2) antibody were assessed atscreening.

A urine drug screen was performed at screening and on Day −1 foralcohol, amphetamines, barbiturates, benzodiazepines, cocainemetabolites, cotinine, methylenedioxymethamphetamine, opiates,phencyclidine, propoxyphene, and tetrahydrocannabinol.

Abnormal clinical laboratory values were flagged as either high or low(or normal or abnormal) based on the reference ranges for eachlaboratory parameter. Clinical significance was defined as any variationin results that had medical relevance and may have resulted in analteration in medical care (e.g., active observation, diagnosticmeasures, or therapeutic measures). If a clinically significant changefrom screening was noted, the clinically significant value and reasonfor clinical significance were documented on the AE page in the eCRF.The subjects were monitored continuously with additional assessmentsuntil the values reached the reference range or the values at screeningor until follow-up was no longer medically necessary.

Vital Sign Measurements

Vital sign measurements included systolic and diastolic blood pressures,heart rate, respiratory rate, and oral body temperature. The subject wasseated for at least 5 minutes before all measurements were taken, withthe exception of orthostatic assessments. At the time points fororthostatic assessments, after taking all measurements with the subjectsseated, subjects were supine for 5 minutes before their blood pressureand heart rate were taken; subjects then stood for 1 minute before theirblood pressure and heart rate were taken again.

Vital signs were measured at the time points indicated in the scheduleof events (Table 1).

Vital sign measurements (systolic and diastolic blood pressures, heartrate, respiratory rate, and oral body temperature) were obtained atscreening; check-in; on Day 1 before the start of infusion and at 1 (endof infusion), 2 and 4 hours (±15 minutes), and 8 and 24 hours (±30minutes) after the start of the infusion; and on Days 3 (±15 minutes forup to 48 hours postdose), 5, 14, 29, 85, and 140. During the infusion,vital sign measurements were obtained every 15 minutes (±5 minutes).Orthostatic assessments were performed at check-in; on Day 1 before thestart of infusion and at 2, 4, 8, and 24 hours (±15 minutes) after thestart of the infusion; and on Day 5.

Clinical significance was defined as any variation in results that hadmedical relevance and may have resulted in an alteration in medical care(e.g., active observation, diagnostic measures, or therapeuticmeasures). If a clinically significant change from screening was noted,the clinically significant value and reason for clinical significancewas documented on the AE page in the subject's eCRF. The subject can bemonitored on a continuing basis with additional assessments until thevalue reached the reference range or the value at screening or untilfollow-up was no longer medically necessary.

Twelve-Lead Electrocardiogram

Single 12-lead ECGs were obtained after the subject had been in thesupine position for at least 5 minutes at the time points indicated inthe schedule of events (Table 1). Twelve-lead electrocardiograms wereobtained at screening; check-in; on Day 1 within 2 hours before thestart of infusion and at 1 (end of infusion), 8, and 24 hours (±15minutes) after the start of the infusion; and on Days 3 (±15 minutes forup to 48 hours postdose), 5, 14, and 140.

Electrocardiogram assessments included comments on whether the tracingswere normal or abnormal, as well as the rhythm, presence of arrhythmiaor conduction defects, morphology, evidence of myocardial infarction,and ST segment, T wave, and U wave abnormalities. In addition,measurements of the following intervals were measured and reported: RRinterval, PR interval, QRS width, QT interval, and QTcF.

Clinical significance was defined as any variation in results that hadmedical relevance and may have resulted in an alteration in medical care(e.g., active observation, diagnostic measures, or therapeuticmeasures). If a clinically significant change from screening was noted,the clinically significant value and reason for clinical significancewere documented on the AE page in the subject's eCRF. The subject wasmonitored continuously with additional assessments until either thevalues reached the reference range or the values at screening or untilfollow up is no longer medically necessary.

Cardiac Telemetry

Cardiac telemetry was performed at the time points indicated in theschedule of events (Table 1). Cardiac telemetry monitoring began on Day1 (starting approximately 30 minutes before the start of infusion) andcontinued for 4 hours after the end of the infusion.

Clinical significance was defined as any variation in results that hadmedical relevance and may have resulted in an alteration in medical care(e.g., active observation, diagnostic measures, or therapeuticmeasures). If a clinically significant change from the initial telemetryfindings on Day 1 (approximately 30 minutes before the start ofinfusion) was noted, the clinically significant value and reason forclinical significance were documented on the AE page in the subject'seCRF. The subject was monitored continuously with additional assessmentsuntil follow-up was no longer medically necessary.

Physical Examination

Complete physical examinations and brief physical examinations wereperformed at the time points indicated in the schedule of events (Table1). A complete physical examination (including an assessment forcutaneous erythema) was performed at screening and on Days 3, 14, and140. A brief physical examination (including an assessment for cutaneouserythema) was performed at check-in (Day −1) and on Days 5, 29, and 85.The physical examination included height and weight at screening andweight only on other days.

A complete physical examination included assessments of the skin(including any signs of cutaneous erythema), head, ears, eyes, nose,throat, neck, thyroid, lungs, heart, cardiovascular system, abdomen,lymph nodes, and musculoskeletal system/extremities. Interim physicalexaminations were to be performed to evaluate AEs or clinical laboratoryabnormalities.

A brief physical examination included assessments of the skin (includingany signs of cutaneous erythema), lungs, cardiovascular system, andabdomen (liver, spleen).

Height and weight were measured and body mass index was calculated onlyat screening. Weight was measured at all other physical examination timepoints indicated in the schedule of events.

Dose Selection for Human Studies

The starting dose in this Phase 1, FIH study was 0.003 mg/kg, which wasdetermined using the NOAEL from a 4-week, repeat-dose toxicology studyin nonhuman primates, the closest and most relevant model to humans andapplying an appropriate safety factor as well as from calculation of theminimum anticipated biological effect level (MABEL) for FB825, based onthe in vitro and in vivo pharmacological data and toxicokinetic data.

The NOAEL for FB825 in the 4-week, repeat dose toxicology study incynomolgus monkeys was considered to be 100 mg/kg. The equivalent humandose, calculated according to the Food and Drug Administration (FDA)Guidance for Industry: Estimating the Maximum Safe Starting Dose inInitial Clinical Trials for Therapeutics in Adult Healthy Volunteers(July 2005), is 38.8 mg/kg; after applying an appropriate safety factorto this (i.e., 100-fold), the maximum recommended starting dose in humansubjects would be approximately 0.39 mg/kg.

The MABEL for FB825 was estimated to be 0.1 μg/mL. It was estimated thata single IV dose in human subjects of 0.003 mg/kg would result insystemic exposure to FB825 in the range from 0.06μg/mL to 0.09μg/mL;thus, the MABEL should not be exceeded during the study. Taking thehigher figure of this range, the expected maximum circulatingconcentration of FB825 with this starting dose would be approximately4.4×10⁴-fold less than the C_(max) for FB825 at the NOAEL (100mg/kg/day) dose level in cynomolgus monkeys, which were administered asingle IV dose of FB825 (Study 20031008). This provides a safety marginof 3.7×10⁴-fold on a milligram per kilogram basis, over and above thehuman equivalent dose of 111.6 mg/kg at the NOAEL (300 mg/kg).

Results

This study evaluated the safety, tolerability, pharmacokinetics, andimmunogenicity of single ascending doses of FB825 via IV injection in arandomized, placebo-controlled, double-blind study in healthy adulthuman subjects. The design and choice of the study population of theplanned first-in-human (FIH) clinical Phase 1 study was based on theneed to obtain initial safety, tolerability, PK, and immunologicaloutcomes for FB825 for future clinical studies. Data was obtainedaccording to the schedule of events presented in Table 1.

FB825 was Safely Administered to Human Subjects

Single, 1 hour, IV infusions of FB825 at doses of 0.003. 0.03, 0.3, 1.5,5, and 10 mg/kg were safe and well tolerated by the healthy subjects inthe clinical trial. There were no deaths and no subject discontinued dueto a treatment emergent adverse event (TEAE). With the exception ofdecreased hemoglobin, upper respiratory tract infection, urinary tractinfection, and gunshot wound, all TEAEs resolved by the end of thestudy.

There were no apparent treatment- or dose-related trends in clinicallaboratory test results, vital sign measurements, 12-lead ECG results,or physical examination findings.

Total IgE Reduced by Administration of FB825

The total IgE was reduced at all postdose time points in the 1.5 and 10mg/kg FB825 treatment groups. In the other doses (0.003, 0.03, 0.3, and5 mg/kg FB825) and placebo treatment groups, the total IgE was reducedat some postdose time points but there were no overall trends (FIG. 1and FIG. 2 ).

Only 4 subjects (1 each in the 0.03, 0.3, and 5 mg/kg FB825 and placebotreatment groups) had detectable ADA, where only 1 subject in the 0.03mg/kg FB825 treatment group had reactive ADA (FIG. 3 ).

Example 3: An Open-Labeled Exploratory Study to Evaluate Safety andEfficacy of FB825 in Adults with Atopic Dermatitis

The instant study is designed to evaluate the change from baseline intotal IgE and allergen-specific IgE in patients having atopic dermatitisafter IV administration of FB825 and to evaluate the clinical efficacyof these patients. This study also aims at evaluating the safety ofFB825 in patients treated thereby, monitoring the changes in clinicalhematology after the IV administration of FB825, and exploring changesfrom baseline in biomarkers, including thymus and activation regulatedchemokine (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP),periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31, and M-CSF after theIV administration of FB825.

FB825 is a humanized monoclonal immunoglobulin G1 (IgG1) targeting theCεmX domain on human B lymphocytic cells expressing membrane-bound IgE(mIgE). FB825 can block the biological pathway of IgE synthesis, thusbenefiting treatment of IgE-mediated allergic diseases. FB825 isformulated in an aqueous solution and given to a patient via a 1-hour IVinfusion at 5 mg/kg.

The heavy chain (top) and light chain (bottom) amino acid sequence ofFB825 in full-length format is provided below (including N-terminalsignal peptide sequences). The heavy chain and light chain CDRs of FB825(following the Kabat numbering scheme) are provided above in FIG. 4 .

(SEQ ID NO: 11) MEFGLSWLFLVAILKGVQCQVQLQESGPGLVKPSETLSLTCTVSGYSITSDYAWNWIRQPPGKGLEWIGSISYSGITGYNPSLKSRVTISRDTSKNQFSLKLSSVTAADTAVYYCARMGYDGLAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK(SEQ ID NO: 12) MRVPAQLLGLLLLWLPGARCDIVMTQTPLSLSVTPGQPASISCRSSQSIVHSNGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPPTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Study Procedures

This is an open-labeled exploratory study to evaluate safety andefficacy of FB825 in adults with atopic dermatitis (AD). Approximately12 human patients with atopic dermatitis (AD), who meet the criteria forstudy entry, were enrolled to the study. All eligible patients receivedFB825, 5 mg/kg, by 1-hour IV infusion on Day 1 and Day 85 (Table 2).Subjects were hospitalized after receiving FB825 on Day 1 and Day 85 andwere discharged from the hospital next day for safety observation (atleast 12 hours). Patients returned to the study site on Days 8, 15, 29,57, 85, 92, 99, 113, 141, and 169 for the safety and efficacy, andbiomarker evaluation (Table 2).

In some instance, patients may be asked to have site visit at Day 78 forblood sampling. The purpose of the blood sampling is to measure totalIgE in order to determine if subjects need the second dose of FB825.Subjects with change from baseline in total IgE less than 50% at Day 78may received the second dose of FB825 (5 mg/kg). Subjects with changefrom baseline in total IgE 50% at Day 78 may have the End of study visitat Day 85 and then complete the study For patients who receive thesecond dose of FB825, the data collected for the second dose may beanalysis and summarized as same as those collected for the first dose.In some instances, patients received the second dose of FB825, 5 mg/kg,by 1-hour IV infusion on Day 85. Such patients were hospitalized beforereceiving FB825 on Day 85 and were discharged from the hospital next dayfor safety observation (at least 12 hours). The patients returned to thestudy site on Days 92, 99, 113, 141 and 169 for the safety and efficacyevaluation.

Serum total IgE and antigen-specific IgE were measured at scheduledvisits and evaluated to explore the changes from baseline after the IVadministration of 5 mg/kg FB825.

Patients were examined for clinical efficacy evaluation activities atscheduled visits, including Pruritus VisualAnalogue Scale (VAS),EczemaArea and Severity Index (EASI), Severity Scoring of AtopicDermatitis Index (SCORAD), Investigator GlobalAssessment (IGA) for ADand Body Surface Area (BSA) involved in AD symptoms.

Safety data, including AEs and laboratory tests, were reviewed by thePI. The duration of subject participation the study, was approximately24 weeks.

Selection of Study Population

Male or female subjects with atopic dermatitis were enrolled in a singlestudy site. Approximately 15 subjects in total were enrolled to achieveat least 12 evaluable subjects.

Selection Criteria (i) Main Inclusion Criteria

Male or female subjects between 20 and 65 years of age, inclusive.

-   -   The subject has a physician-confirmed diagnosis of chronic        atopic dermatitis based on 3 years history of symptoms defined        by the Eichenfield revised criteria of Hannifin and Rajka and        supported by positive allergen-specific IgE at the screening        visit.    -   Eczema Area and Severity Index (EASI) score ≥14 at the screening        and baseline visits.    -   Investigator's Global Assessment (IGA) score ≥3 (5-point scale)        at the screening and baseline visits.    -   ≥10% body surface area (BSA) of AD involvement at the screening        and baseline visits.    -   History of inadequate response to a stable (1 month) regimen of        topical corticosteroids or calcineurin inhibitors as treatment        for AD within 3 months before the screening visit. (The regimen        of topical corticosteroids means medium to high potency, applied        for at least 28 days or for the maximum duration recommended by        product prescribing information.)    -   Patients must be applying stable doses of emollient provided for        atopic dermatitis twice-daily for at least 7 days before the        baseline visit.    -   Female subjects of childbearing potential must use at least two        forms of birth control. One must be barrier protection (i.e.,        condom or female condom) and the other is one of acceptable        method of birth control (i.e., diaphragm, intrauterine device,        hormonal contraceptives, or abstinence) throughout the study.        Subjects who are surgically sterile (i.e., hysterectomy,        bilateral tubal ligation, or bilateral oophorectomy), or        postmenopausal (defined as amenorrhea for 12 consecutive months        and documented serum follicle stimulating hormone level >40        mU/mL) will be considered as no childbearing potential. All        female subjects must have a negative serum pregnancy test prior        to dosing. The subject must use the method of contraception        mentioned above during the study period and in 16 weeks or 5        half-lives after the last dosing of FB825.    -   The subject has a body weight ≥40 kg at screening and a body        mass index of 18.0 to 30.0 kg/m², inclusive.    -   The subject has a normal, as determined by the investigator,        12-lead electrocardiogram (ECG) with normal cardiac conduction        parameters:        -   Heart rate between 45 and 100 bpm;        -   Fridericia-corrected QT interval (QTcF) ≤450 milliseconds            (men) or ≤470 milliseconds (women); and        -   QRS interval lower than 120 milliseconds.    -   The subject is healthy, except atopic diseases, as determined by        the investigator, on the basis of clinical laboratory test        results performed at screening. If the results are outside the        normal reference ranges, the subject may be included only if the        investigator judges the abnormalities or deviations from normal        not to be clinically significant. This determination must be        recorded in the subject's source document and initialed by the        investigator. This is not applicable to the laboratory        abnormalities listed in the exclusion criterion (using the        Division of Microbiology and Infectious Diseases criteria).    -   The subject is able to provide written informed consent.    -   The subject agrees to comply with all protocol requirements.

(ii) Main Exclusion Criteria

-   -   Female subjects who are pregnant or lactating.    -   The subject is on diet or with poor intake.    -   The subject has a history of heart arrhythmias (any clinically        relevant).    -   The subject has a positive test result for hepatitis B surface        antigen, hepatitis C virus antibody, or human immunodeficiency        virus antibodies at screening.    -   The subject has a history of alcohol or drug abuse that would        impair or risk the patients' full participation in the study, in        the opinion of the investigator.    -   The subject is under judicial supervision or curatorship.    -   The subject has a clinically relevant, currently active or        underlying gastrointestinal cardiovascular, nervous system,        psychiatric, metabolic, renal, hepatic, respiratory (with the        exception of uncomplicated allergic rhinitis), inflammatory,        immunological, endocrine, diabetes, or infectious disease and        ineligible to participate in the study judged by investigator.    -   The subject has any history of a previous anaphylactic reaction.    -   The subject has any condition that, in the opinion of the        investigator, would compromise the study or the well-being of        the subject or prevent the subject from meeting or performing        study requirements.    -   The subject has received any immunoglobulin products or blood        products within 3 months prior to dosing.    -   The subject has received a biologic product:        -   The subject has received any cell-depleting agents, not only            limited to rituximab, within 6 months prior to dosing, or            before the lymphocyte count returns to normal, whichever is            longer.        -   The subject has received other biologics within 5 half-lives            (if known) or 16 weeks, which is longer, prior to dosing).    -   The subject has one or more of the following laboratory        abnormalities at screening as defined by Division of        Microbiology and Infectious Diseases Adult Toxicity Table, 2007        [Laboratory values may be converted to equivalent standard        units. Retesting of abnormal laboratory values that may lead to        exclusion will be allowed once (without prior sponsor approval).        Retesting will take place during an unscheduled visit in the        screening phase (before baseline)]:        -   Aspartate aminotransferase or alanine aminotransferase            (>2×upper limit of normal [ULN]) or higher;        -   Total bilirubin ≥1.5×ULN        -   Serum creatinine ≥1.6×ULN        -   Any other laboratory abnormality higher than or equal to            grade 2 with the exception of IgE level, eosinophil counts,            eosinophil cationic protein (ECP) and laboratory values            mentioned above.

Laboratory values may be converted to equivalent standard units.Retesting of abnormal laboratory values that may lead to exclusion maybe allowed once (without prior sponsor approval). Retesting may takeplace during an unscheduled visit in the screening phase (beforebaseline).

-   -   The subject has received any approved or unapproved (i.e.,        investigational) immunotherapy treatment within the past 3        months.    -   The subject has used any of the following classes of medication        (prescription or over the counter):        -   Intranasal corticosteroid (e.g., fluticasone propionate)            within 30 days prior to dosing.        -   Systemic corticosteroids (e.g., prednisone) within 30 days            prior to dosing.        -   Leukotriene modifiers (e.g., montelukast) within 30 days            prior to dosing.        -   Immunosuppressants (e.g., gold salts, methotrexate,            azathioprine, cyclosporine) within the past 30 days prior to            dosing.        -   Immunomodulating drugs (e.g., IFN-γ) within the past 30 days            prior to dosing.        -   Anti-IgE (e.g., omalizumab) within the past 1 year prior to            dosing.        -   Allergen immunotherapy within the past 1 year prior to            dosing.        -   Orally inhaled corticosteroids (e.g., budesonide) within the            past 30 days prior to dosing.    -   The subject has received phototherapy within 4 weeks prior to        dosing.    -   The subject has received live vaccine within 12 weeks prior to        dosing.    -   The subject has known or suspected history of immunosuppression,        including history of opportunistic infections (e.g., TB) per        investigator judgment.    -   The subject has history of malignancy within 5 years before the        screening period.    -   High risk of parasite infection. Risk factors for parasitic        disease (living in an endemic area, chronic gastrointestinal        symptoms, travel within the last 6 months to regions where        geohelminthic infections are endemic, and/or chronic        immunosuppression) AND Evidence of parasitic colonization or        infection on stool evaluation for ova and parasites. Stool ova        and parasite evaluation will only be conducted in patients with        risk factors and an eosinophil count more than twice the upper        limit of normal subjects.

Study Treatments

(i) Dosing and IV Administration of Study Treatment

Two doses of 5 mg/kg FB825 was given to subjects with atopic dermatitis.Subjects received FB825 by 1-hour IV infusion in the morning at Day 1and Day 85. Fasting for at least 2 hours was required in some instance.Water intake is not allowed within 1 hour prior to dosing and 1 hourafter dosing. The administered amount of FB825 can be adjusted based onsubject's body weight, the appropriate amount of drug product wasdiluted with 250 mL 0.9% sodium chloride solution. FB825 wasadministered via IV route over 1 hour with the aid of a programmablevolumetric infusion device. The final diluted product with 0.9% sodiumchloride, after reconstitution, has to be used as soon as possible, andhas to be used with 8 hours. The reconstituted FB825 can be stored at 2°C. to 25° C. for a maximum of 8 hours, prior to use.

(ii) Prior, Concomitant and Prohibited Medications

Prior Medications and Therapies

Information about prior medication taken by the subject within the 30days before he or she provides informed consent was recorded in thesubject's CRF.

Pre-Treatment/Concomitant Medication and Procedures

Any treatment (including nutritional supplements) or procedureadministered from the time of signing of the ICF to the end of studyvisit is considered concomitant and was recorded in the CRF. Thisincludes permitted medications ongoing at the time of consent.

The AD basal therapy during the study described below:

-   -   All patients are required to apply moisturizers at least twice        daily for at least the 7 consecutive days prior to dosing and to        continue the treatment throughout the study. All types of        moisturizers are permitted, but patients may not initiate        treatment with prescription moisturizers or moisturizers        containing additives during the screening period or during the        study.    -   Patients may continue using stable doses of prescription        moisturizers or moisturizers containing additives, if initiated        before the screening visit. Starting on day 1/baseline, all        patients are required to initiate treatment with topical        corticosteroid (TCS) using a standardized regimen according to        the following guidelines:    -   Apply medium potency TCS daily to areas with active lesions. Low        potency TCS should be used on areas of thin skin (face, neck,        intertriginous, and genital areas, areas of skin atrophy, etc.)        or for areas where continued treatment with medium potency TCS        is considered unsafe.    -   After lesions are under control (clear or almost clear), switch        from medium potency to low potency TCS and treat daily for 7        days, then stop.    -   If lesions return, reinstitute treatment with medium potency        TCS, with the step-down approach described above upon lesion        resolution.    -   For lesions persisting or worsening under daily treatment with        medium potency TCS, patients may be treated (rescued) with high        or super-high potency TCS, unless higher potency TCS are        considered unsafe.    -   Monitor the patient for signs of local or systemic TCS toxicity        and step down or stop treatment as necessary.    -   The type and amount of topical products used during the study        were recorded. The amount of TCS used was determined by weighing        the tube at each visit (see study reference manual for details).    -   It is recommended that patients use fluticasone propionate 0.05%        cream, mometasone furoate 0.1% cream, or betamethasone valerate        0.06% cream as medium potency TCS, and hydrocortisone 1%        ointment for low potency TCS.    -   If rescue with TCS is needed, it is recommended that patients        use fluocinonide 0.05% cream, desoximetasone 0.25% ointment as        high potency TCS, and clobetasol propionate 0.05% ointment for        super high potency TCS.    -   Do not use moisturizers and TCS on the same areas at the same        time during the day. On areas not treated with TCS, moisturizers        will be applied twice daily—morning and evening.

Pre-treatment medication/procedures: medications taken or proceduresperformed prior to dosing.

Concomitant medication/procedures: medications taken or proceduresperformed following the IV administration of study drug through the EOSvisit.

Prohibited Concomitant Medications:

-   -   Tropical tacrolimus and pimecrolimus    -   Systemic corticosteroids, unless the subjects are under rescue        medication.    -   Leukotriene inhibitors    -   Allergen immunotherapy    -   Systemic treatment for AD with an        immunosuppressive/immunomodulating substance (including, but not        limited to, cyclosporine, mycophenolate-mofetil, IFN-γ,        azathioprine, methotrexate, or biologics)    -   Treatment with a live (attenuated) vaccine    -   Traditional Chinese Medicine

Prohibited Concomitant Procedures:

-   -   Surgical procedures    -   Concomitant ultraviolet (UV) procedures (phototherapy [NBUVB,        UVB, UVA1, or PUVA])    -   Tanning in a bed/booth is not allowed during the study    -   Patients are not allowed more than 2 bleach baths per week        during study participation

(iii) Handling of Infusion-Related or Allergic Reaction

The IV administration of study drug must be performed under supervisionof trained medical staff and where facilities to handle allergicreactions are available. If a subject experiences an infusion-relatedreaction, the subject must be treated symptomatically with supportivecare, further monitoring, and appropriate medical therapy which mayinclude antihistamines and/or corticosteroid if needed. The studyinfusion may be stopped and the subject would followed until the end ofthe study. The amount infused was recorded. Should a subject experiencesymptoms typical of an allergic reaction (eg, shortness of breath,anaphylaxis, urticaria, angioedema), then study drug IV administrationshould be discontinued immediately and permanently.

Suspected anaphylaxis should be assessed according to the clinicaldiagnostic criteria outlined by the National Institute of Allergy andInfectious Diseases which are provided in Appendix 12-2.

For these and other circumstances, subjects may receive appropriatemedical treatment at the discretion of the investigator.

In case of allergic reactions, patients may be rescued with a prohibitedmedication or procedure to treat intolerable AD symptoms.

-   -   If medically necessary (i.e., to control intolerable AD        symptoms), rescue treatment with systemic corticosteroids for AD        at doss less than 1 mg/kg/day prednisolone and no more than 3        days may be provided to study patients after week 2.    -   Patients were subject to efficacy and safety assessments (e.g.,        disease severity scores, safety labs) immediately before        administering any rescue treatment.

Study Procedures and Methods of Assessment

The following sections describe the study procedures and data to becollected. Subjects were assessed by the same investigator or sitepersonnel whenever possible. The schedule and assessment is provided inTable 2. The baseline characteristics are provided in Table 3.

(i) Endpoints:

Primary Endpoint(s):

-   -   Change from baseline in total IgE at Day85 and day 169/End of        Study (EOS).    -   Change from baseline in allergen-specific IgE at Day 85 and Day        169/EOS.

Endpoints for Biomarker:

-   -   Change from baseline in total IgE at Day 8, 15, 29, 57, 85, 92,        99, 113, 141 and 169.    -   Change from baseline in allergen-specific IgE at Day 8, 15, 29        and 57, 85, 92, 99, 113, 141, and 169. Change from baseline in        biomarkers including thymus and activation regulated chemokine        (TARC), Eotaxin-3, thymic stromal lymphopoietin (TSLP),        periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31 and M-CSF        after IV administration of FB825 at Days 8, 15, 29, 57, 85, 92,        99, 113, 141, and 169.

Efficacy Endpoints:

-   -   Changes from baseline in Pruritus Visual Analogue Scale (VAS) at        Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.    -   Changes from baseline in Eczema Area and Severity Index (EASI)        at Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.    -   Changes from baseline in Severity Scoring of Atopic Dermatitis        Index (SCORAD) at Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and        169.    -   Changes from baseline in Investigator Global Assessment (IGA)        for atopic dermatitis at Days 8, 15, 29, 57, 85, 92, 99, 113,        141, and 169.    -   Changes from baseline in Body Surface Area (BSA) involved in        atopic dermatitis at Days 8, 15, 29, 57, 85, 92, 99, 113, 141,        and 169.    -   Change from baseline in biomarkers including thymus and        activation regulated chemokine (TARC), Eotaxin-3, thymic stromal        lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13,        IL-16, IL-31 and M-CSF after IV administration of FB825 at Days        8, 15, 29, 57, 85, 92, 99, 113, 141, and 169.    -   Safety was assessed by monitoring and recording of adverse        events (AEs) and serious adverse event (SAEs); physical        examination findings and vital sign measurements (systolic and        diastolic blood pressures, heart rate, respiratory rate, and        body temperature), clinical laboratory test results (hematology,        coagulation, serum chemistry [including liver function tests,        blood glucose level], and urinalysis); 12-lead ECG results.

(ii) Biomarker Assessments

Blood samples for the measurement of total, allergen-specific IgE,thymus and activation regulated chemokine (TARC), Eotaxin-3, thymicstromal lymphopoietin (TSLP), periostin, IL-1a, IL-4, IL-5, IL-13,IL-16, IL-31 and M-CSF was analyzed according to methodology describedin a separate report.

Blood samples was collected. The actual sample collection time andsampling problems, if any, was recorded on the CRF. Approximately 5 mLsample of venous blood per sample was drawn at the following timepoints:

Total Immunoglobulin/Allergen-specific IgE:

-   -   Screening period: at any time    -   Day 1, and 85: (In 2 hours before the start of infusion)    -   Days 8, 15, 29, 57, 92, 99, 113, and 141, and 169 after IV        administration of FB825 at any time on the day.

TARC, Eotaxin-3, TSLP, periostin, IL-1a, IL-4, IL-5, IL-13, IL-16, IL-31and M-CSF:

-   -   Day 1 and 85 in 2 hours before the start of infusion, and at any        time on Days 8, 15, 29, 57, 92, 99, 113, 141, and 169.

(iii) Clinical Efficacy Assessments

Pruritus Visual Analogue Scale (VAS).

The pruritus visual analogue scale (VAS) of SCORAD was applied for themeasurement of pruritus. Patients were asked to assign a numerical scorerepresenting the intensity of their symptoms on a scale from 0 to 10,with 0 for having no symptoms and 10 having worst symptoms. Subjectswere asked to perform the measurement at screening, Day 1 and 85 beforedosing, Day 2 and Day 86 before discharge, and any time of Day 8, 15,29, 57, 92, 99, 113, 141, and 169.

Severity Scoring of Atopic Dermatitis Index (SCORAD)

The SCORAD (Index) is the validated scoring system in atopic dermatitis(AD). To measure the extent of AD, the rule of nines is applied on afront/back drawing of the patient's inflammatory lesions. The extent canbe graded from 0 to 100. The intensity part of the SCORAD consists of 6items: erythema, oedema/papulation, excoriations, lichenification,oozing/crusts and dryness. Each item can be graded on a scale from 0(absent) to 3 (severe). The subjective items include daily pruritus andsleeplessness. The SCORAD Index formula is: A/5+7B/2+C. In this formulaA is defined as the extent (0-100), B is defined as the intensity (0-18)and C is defined as the subjective symptoms (0-20). The maximal score ofthe SCORAD Index is 103. Subjects were asked to perform the measurementat screening, Day 1, and 85 before dosing, Day 2 and Day 86 beforedischarge, and any time of Day 8, 15, 29, 57, 92, 99, 113, 141, and 169.

Eczema Area and Severity Index (EASI)

The EASI scoring system uses a defined process to grade the severity ofthe signs of eczema and the extent affected. Extent and severity ofsigns of eczema was evaluated in four body regions and the total scoreis the sum of the four regions scores adjusted with multipliers. TheEASI score is ranged from 0-72. Subjects were asked to perform themeasurement at screening, Day 1 and 85 before dosing, Day 2 and Day 86before discharge, and any time of Day 8, 15, 29, 57, 92, 99, 113, 141,and 169.

Investigator Global Assessment (IGA) for AD

IGA allows investigators to assess overall disease severity at one giventime point, and it consists of a 5-point severity scale from clear tovery severe disease (0=clear, 1=almost clear, 2=mild disease, 3=moderatedisease, and 4=severe disease). Subjects were asked to perform themeasurement at screening, Day 1 and 85 before dosing, Day 2 and Day 86before discharge, and any time of Day 8, 15, 29, 57, 92, 99, 113, 141,and 169.

BSA Involved in AD Symptoms

It was measured as part A (Extent) of SCORAD. Subjects were asked toperform the measurement at screening, Day 1 and 85 before dosing, Day 2and Day 86 before discharge, and any time of Day 8, 15, 29, 57, 92, 99,113, 141, and 169.

(iv) Safety Assessments

Safety was assessed by monitoring and recording of Adverse Event (AEs),Serious Adverse Event (SAEs), physical examination findings and vitalsign measurements (systolic and diastolic blood pressures, heart rate,respiratory rate, and oral body temperature), clinical laboratory testresults (hematology, coagulation, serum chemistry [including liverfunction tests], and urinalysis), and 12-lead ECG results. The overallsummary of numbers of patients with adverse events through the 15-168days treatment period-SAF is provided in Table 4.

Adverse Events

The Adverse Event section (Section 7) describes the Adverse Event(SAEs), Serious Adverse Event (SAEs) and Adverse Events of SpecialInterest were collected during the study.

Physical Examinations

A complete physical examination was performed at the time pointsindicated in the schedule of events.

A complete physical examination includes assessment of skin (includingany signs for cutaneous erythema), head, ears, eyes, nose, throat, neck,thyroid, lungs, heart, cardiovascular system, abdomen, lymph nodes, andmusculoskeletal system/extremities. Interim physical examinations wereperformed at the discretion of the investigator, if necessary, toevaluate AEs or clinical laboratory abnormalities. Height and weight wasmeasured and body mass index will be calculated at screening only.Weight was also measured at all other physical examination time pointsas indicated in the schedules of events for the study. Body weight wasrecorded in kilograms (kg) to 1 decimal place in indoor clothing(without coat and shoes) and body height (without shoes) was measured incentimeters (cm) without decimal places.

Vital Sign Measurements

Vital sign measurements include systolic and diastolic blood pressures,heart rate, respiratory rate, and body temperature. The subject wasseated for at least 5 minutes before all measurements are taken. Vitalsigns were measured at the time points indicated in the schedule ofevents.

When procedures are overlapping and occurring at the same time point,the order of procedures should be vital sign measurements and then ECGs.

The investigator may determine whether any of the vital signmeasurements are clinically significant or not clinically significant.Clinical significance is defined as any variation in results that hasmedical relevance and may result in an alteration in medical care (e.g.,active observation, diagnostic measures, or therapeutic measures). If aclinically significant change from the screening values is noted, theclinically significant value and reason for clinical significance wasdocumented in the AE page of the subject's CRF. The investigator maycontinue to monitor the subject with additional assessments until thevalue has reached the reference range or the value at screening or untilthe investigator determines that follow-up is no longer medicallynecessary.

Clinical Laboratory Testing

Clinical laboratory tests were performed by local site. Blood and urinewere be collected under fasting conditions (fasted for approximately 2or more hours) at the time points indicated in the schedule of events.

The following hematology, coagulation, serum chemistry (including liverfunction and thyroid function tests), urinalysis assessments wereperformed:

-   -   Hematology: Hematocrit (Hct), hemoglobin (Hb), mean corpuscular        hemoglobin (MCH),-mean corpuscular hemoglobin concentration        (MCHC), mean corpuscular volume (MCV), platelet count, red blood        cell (RBC) count, and WBC and differential count (absolute and        percent).    -   Coagulation: International normalized ratio (INR), partial        thromboplastin time (PTT), and -prothrombin time (PT)    -   Serum Chemistry: ALT, albumin, alkaline phosphatase, amylase,        anion gap, AST, bicarbonate, bilirubin (total and direct), blood        urea nitrogen (BUN), calcium, carbon dioxide, chloride,        cholesterol (total, high-density lipoprotein, and calculated        low-density lipoprotein), creatine phosphokinase, creatinine,        gamma-glutamyltransferase (γ-GT), globulin, glucose, lactate        dehydrogenase (LDH), lipase, magnesium, phosphorus, potassium,        sodium, total protein, triglycerides, troponin I or T, and uric        acid.    -   Thyroid Function: Free T4 and thyroid-stimulating hormone    -   Urinalysis: Appearance, bilirubin, color, glucose, ketones,        leukocyte esterase, microscopy (performed if dipstick is        positive; includes bacteria, casts, crystals, epithelial cells,        red blood cells, and white blood cells), nitrites, occult blood,        pH, protein, specific gravity, turbidity, and urobilinogen

A serum pregnancy test (β-human chorionic gonadotropin) will beperformed on all female subjects with potential of children bearing atscreening and confirmed result prior to dosing. Urine pregnancy testswere performed on Days 8, 15, 29, 57, 85, 92, 99, 113, 141, and 169. Forfemale subjects who were postmenopausal, a serum follicle-stimulatinghormone test was performed at screening.

Hepatitis B surface antigen, hepatitis C virus antibody, and humanimmunodeficiency virus antibody will be assessed at screening.

Abnormal clinical laboratory values were flagged as either high or low(or normal or abnormal) based on the reference ranges for eachlaboratory parameter. The investigator may determine whether any of theabnormally high or low results are clinically significant or notclinically significant. Clinical significance is defined as anyvariation in results that has medical relevance and may result in analteration in medical care (e.g., active observation, diagnosticmeasures, or therapeutic measures). If a clinically significant changefrom the screening value is noted, the clinically significant value andreason for clinical significance may be documented in the AE page of theCRF. The investigator may continue to monitor the subject withadditional assessments until the values have reached the reference rangeor the values at screening or until the investigator determines thatfollow-up is no longer medically necessary.

Clinically significant laboratory values for individual subjects werelisted. A summary for the number and percentage of subjects withclinically significant laboratory values at any time point waspresented.

Electrocardiograms

Single 12-lead ECGs were obtained after the subject has been in thesupine position for at least 5 minutes at the time points indicated inthe schedule of events. Electrocardiogram assessments include commentson whether the tracings are normal or abnormal, rhythm, presence ofarrhythmia or conduction defects, morphology, any evidence of myocardialinfarction, or ST segment, T wave, and U wave abnormalities. Inaddition, measurements of the following intervals will be measured andreported: RR interval, PR interval, QRS width, QT interval, and QTcF.

Adverse Events

The overall summary of number of patients with adverse events throughthe 15-168 days treatment period-SAF is provided in Table 4.

(i) Definitions Adverse Event (AE)

An AE is defined as any untoward medical occurrence associated with theuse of a drug in humans, whether or not considered drug related.Subjects will be instructed to contact the investigator at any timeduring the study period if any symptoms develop.

A TEAE is defined as any event not present before exposure to study drugor any event already present that worsens in intensity or frequencyafter exposure.

An adverse reaction is any AE caused by a drug. Adverse reactions are asubset of all suspected adverse reactions for which there are reasons toconclude that the drug caused the event.

A suspected adverse reaction is any AE for which there is a reasonablepossibility that the study drug caused the AE. For the purposes ofinvestigational new drug safety reporting, “reasonable possibility”means that there is evidence to suggest a causal relationship betweenthe study drug and the AE. A suspected adverse reaction implies a lesserdegree of certainty about causality than adverse reaction, which meansany AE caused by a study drug.

An AE or suspected adverse reaction is considered “unexpected” if it isnot listed in the IB or at the specificity or severity that has beenobserved with the study drug being tested; or, if an IB is not requiredor available, is not consistent with the risk information described inthe general investigational plan or elsewhere in the currentapplication. For example, under this definition, hepatic necrosis wouldbe unexpected (by virtue of greater severity) if the IB referred only toelevated hepatic enzymes or hepatitis. Similarly, cerebralthromboembolism and cerebral vasculitis would be unexpected (by virtueof greater specificity) if the IB listed only cerebral vascularaccidents. “Unexpected,” as used in this definition, also refers to AEsor suspected adverse reactions that are mentioned in the IB as occurringwith a class of drugs or as anticipated from the pharmacologicalproperties of the drug, but are not specifically mentioned as occurringwith the particular drug under investigation.

Serious Adverse Event (SAE)

An AE or suspected adverse reaction is considered an SAE if, in the viewof either the investigator or sponsor, it results in any of thefollowing outcomes:

-   -   Death    -   Life-threatening AE    -   Inpatient hospitalization or prolongation of existing        hospitalization    -   Persistent or significant incapacity or substantial disruption        of the ability to conduct normal-life functions    -   Congenital anomaly or birth defect

Important medical events that may not result in death, be lifethreatening, or require hospitalization may be considered serious when,based upon appropriate medical judgment, they may jeopardize the subjectand may require medical or surgical intervention to prevent one of theoutcomes listed in this definition. Examples of such medical eventsinclude allergic bronchospasm requiring intensive treatment in anemergency room or at home, blood dyscrasias or convulsions that do notresult in inpatient hospitalization, or the development of drugdependency or drug abuse.

An AE or suspected adverse reaction is considered “life threatening” if,in the view of either the investigator or sponsor, its occurrence placesthe subject at immediate risk of death. It does not include an AE orsuspected adverse reaction that, had it occurred in a more severe form,might have caused death.

Adverse Events of Special Interest

Adverse events of special interest include the following events:

-   -   Any subject experiences a treatment-emergent AE (TEAE) of        anaphylaxis    -   Any subject experiences an SAE (Section 7.1.2)    -   Any subject experiences a persistent QT prolongation (>500        milliseconds, or ≥60 milliseconds change from baseline) for at        least 30 minutes or ischemic changes on repeated ECGs, or        persistent symptomatic arrhythmia    -   Hypersensitivity reactions including anaphylaxis, thyroid        abnormalities, cutaneous erythema, thrombocytopenia, anemia,        hemolysis, neutropenia, hepatotoxicity, and nephrotoxicity.    -   The following laboratory parameters are encountered in any        individual subject:        -   ALT or AST ≥5×ULN    -   Bilirubin ≥2×ULN    -   Platelet count grade 1≤99.999*109/L    -   Hemoglobin ≤105 g/L    -   Absolute neutrophil count grade 1≤1.5*109/L    -   Blood urea nitrogen or creatinine rise to >2×ULN

(ii) Eliciting of Adverse Events

The investigator is responsible for ensuring that all AEs and SAEs arerecorded in the CRF and reported to Fountain. Adverse events wereassessed from the time of screening period until completion of all studyprocedures and discharge from the study.

At every study visit or assessment, subjects were asked a standardquestion to elicit any medically-related changes in their well-being.They were also asked if they had been hospitalized, had any accidents,used any new medications, or changed concomitant medication regimens(both prescription and over-the-counter medications).

In addition to subject observations, AEs were documented from any datacollected in the AE page of the CRF (eg, laboratory values, physicalexamination findings, and ECG changes) or other documents that arerelevant to subject safety.

Assessment for Severity The severity (or intensity) of an AE using theCommon Terminology Criteria for Adverse Events (CTCAE) was used forinfusion reaction severity grading.

-   -   Grade 1 Mild: Transient or mild discomfort (<48 hours); no        medical intervention/therapy-required.    -   Grade 2 Moderate: Mild to moderate limitation in activity—some        assistance may be needed; no or minimal medical        intervention/therapy required.    -   Grade 3 Severe: Marked limitation in activity, some assistance        usually required; medical intervention/therapy required,        hospitalization possible.    -   Grade 4 Life threatening: Extreme limitation in activity,        significant assistance required; significant medical        intervention/therapy required, hospitalization or hospice care        probable.

Changes in the severity of an AE should be documented to allow anassessment of the duration of the event at each level of intensity to beperformed. An AE characterized as intermittent requires documentation ofonset and duration of each episode.

7.3.2 Assessment of Causality

The investigator's assessment of an AE's relationship to study drug ispart of the documentation process but is not a factor in determiningwhat is or is not reported in the study.

The investigator assessed causality (i.e., whether there is a reasonablepossibility that the study drug caused the event) for all AEs and SAEs.The relationship was characterized using the following classification:

-   -   Unrelated: This relationship suggests that there is no        association between the study drug and the reported event.    -   Possible: This relationship is based on evidence suggesting a        causal relationship between the study drug and the AE, i.e.,        there is a reasonable possibility that the drug caused the        event. The event follows a reasonable temporal sequence from the        time of drug IV administration or follows a known response        pattern to the study drug, but could also have been produced by        other factors.    -   Probable: This relationship suggests that a reasonable temporal        sequence of the event with drug IV administration exists and,        based upon the known pharmacological action of the drug, known        or previously reported adverse reactions to the drug or class of        drugs, or judgment based on the investigator's clinical        experience, the association of the event with the study drug        seems likely.    -   Definite: This relationship suggests that a definite causal        relationship exists between drug IV administration and the AE,        and other conditions (concurrent illness, progression/expression        of disease state, or concurrent medication reaction) do not        appear to explain the event.

Statistical Analysis

(i) General Statistics

Details of all statistical analyses were described in a statisticalanalysis plan. All data collected was presented in data listings. Datafrom subjects excluded from an analysis population was presented in thedata listings but not included in the calculation of summary statistics.

For categorical variables, frequencies and percentages were presented.Continuous variables were summarized using descriptive statistics(number of subjects, mean, median, SD, minimum, and maximum).

Baseline demographic and background variables were summarized by doseand overall for all subjects. The number of subjects who enroll in thestudy and the number and percentage of subjects who complete the studywas presented. Frequency and percentage of subjects who withdraw ordiscontinue from the study, and the reason for withdrawal ordiscontinuation, was also be summarized. Analysis for this study wasdemonstrated with descriptive statistics for each period and group. Nosignificance test was applied.

A statistical analysis plan (SAP) was written to address statisticalanalysis work in detail. The clinical database lock may occur after alldata are reconciled (i.e., “cleaned”) after the last patient completesthe study.

(ii) Sample Size Calculations

A total of approximately 12 evaluable subjects were planned for thisstudy. The sample size for this study is based on clinical and practicalconsiderations and not on a formal statistical power calculation.

(iii) Analysis Sets

Full Analysis Set (FAS) is defined as subjects who received at least 1dose of study drug. All efficacy evaluation will be performed in FASpopulation.

The Safety population is defined as subjects who received at least 1dose of study drug. All safety evaluation will be performed in safetypopulation.

(iv) Statistical Analysis

Biomarker Analyses

Concentration and change from baseline in total and allergen-specificIgE and biomarkers listed as endpoints was summarized by visit andpresented graphically. Change from baseline in total andallergen-specific IgE was be summarized by the baseline total IgEconcentration (serum IgE>1500 IU/mL or serum IgE ≤1500 IU/mL) and givenFB825 doses (1 dose or 2 doses).

In subjects who receive the second dose of FB825, the data collected forthe second dose were analysis and summarized as same as those collectedfor the first dose.

Clinical Efficacy Analysis

The evaluation index was assessed by PI or co-PI during every visit foreach subject. The clinical endpoints were analyzed using descriptivestatistic (mean of EASI, SCORAD, IGA, VAS, and BSA; SD, CV, number ofsubjects) by visit time points and given FB825 doses (1 dose or 2doses).

The change of mean index versus scheduled visit time profiles waspresented graphically,

In subjects who receive the second dose of FB825, the data collected forthe second dose was analysis and summarized as same as those collectedfor the first dose.

Safety Analyses

Adverse events were coded by preferred term and system organ class usingthe latest version of the Medical Dictionary for Regulatory Activitiesand summarized by treatment, dose level, and overall. Adverse eventswere also summarized by severity, relationship to study drug, SAEs, andAEs leading to discontinuation of study drug.

Actual values and changes from baseline for clinical laboratory testresults, vital sign measurements, and 12-lead ECG results weresummarized by treatment and dose at each time point using descriptivestatistics (number of subjects, mean, SD, median, minimum, and maximum).Shift tables were generated for clinical laboratory test results.Clinical laboratory data, vital sign measurements, 12-lead ECG results,and physical examination findings were presented in data listings.

(v) Handling of Missing Data

Concentrations that are below the limit of quantification (BLQ) weretreated as zero for descriptive statistics. Mean BLQ concentrations willbe presented as BLQ, and the SD and CV were reported as not applicable.Missing concentrations will be excluded from the calculations.

Last observation carry forward (LOCF) was applied to handle missingdata. No imputation will be available for safety data.

(vi) Data Quality Assurance

All aspects of the study were monitored for compliance with applicablegovernment regulations with respect to current International Conferenceon Harmonisation (ICH) harmonized tripartite guideline E6 (R1): GoodClinical Practice and current standard operating procedures. There maybe an internal quality review audit of the data and additional reviewsby the clinical monitor.

TABLE 2 Schedule of Event and Assessment Procedure Dose 1 FB825 Dose 2FB825 Day Day Day Day Day Day Day Day Day Day Screening Day Day 8 15 2957 85^(a) Day 92 99 113 141 169/EOS^(b) Study Day(s) −23 to −1 1^(a) 2(±1) (±1) (±2) (±2) (±3) 86 (±1) (±1) (±2) (±2) (±3) Informed consent XInclusion/ X X Exclusion criteria Medical history X Physical examinationX X X X X X X X X X X X Vital sign X X X X X X X X X X X X X Xmeasurements ^(c,d) 12-Lead X X X X X X X X X X X X Electrocardiogram^(e) AD symptom evaluation ^(f) X X X X X X X X X X X X X X Serology^(g) X Clinical laboratory X X X X X X X X X X X X X X testing ^(h)Thyroid function testing X X X X X X X X Urine drug, and blood X X^(m) XX alcohol test Pregnancy test (female X X X X X X X X X X X X subjects)^(i) Serum FSH ^(j) X Admission to clinic X X Study drug IV X Xadministration Total Immunoglobulin/ X X X X X X X X X X X XAllergen-specific IgE ^(k) Biomarker Assessments ^(l) X X X X X X X X XX X Adverse events assessment X X X X X X X X X X X X X XPrior/Concomitant X X X X X X X X X X X X X X medications Discharge fromclinic X X Abbreviations: ALT, alanine aminotransferase; AST, aspartateaminotransferase; EOS, end of study; FSH, follicle-stimulating hormone;Ig, immunoglobulin. ^(a)Subjects will check into the clinic on Day 1 andDay 85 and have check-in procedures performed. ^(b)Subjects earlywithdraw from the study will be requested to have an EOS visit andperform required activities. ^(c) Vital sign measurements (systolic anddiastolic blood pressures, heart rate, respiratory rate, and bodytemperature) will be obtained at screening; Day 1, and 85 before thestart of infusion and all study visits. Vital sign measurements will beobtained at 1.25 (end of infusion), 2, 4, 8 hours (±5 minutes in eachtime points) after the start of the infusion on Day 1, and Day 85,andprior to discharge on Day 2 and Day 86. During the infusion, vital signmeasurements will be obtained every 15 minutes (±5 minutes). ^(d) Whenprocedures are overlapping and occurring at the same time point, theorder of procedures should be vital sign measurements and then ECGs.^(e) Twelve-lead electrocardiograms will be obtained at screening; onDay 1 and Day 85 within 2 hours before the start of infusion and at 1.25(end of infusion, ±5 minutes) 8 hours (±5 minutes) after the start ofthe infusion; prior to discharge; and all other study visits, except Day57, and 141. ^(f) AD symptom evaluation is determined by VAS, EASI,SCORAD, IGA, and BSA. These are evaluated by Investigator at screening;on Day 1 prior to dosing, and all other study visits. ^(g) Serologytesting will include hepatitis B surface antigen, hepatitis C virusantibody, and human immunodeficiency virus antibodies. ^(h) Clinicallaboratory tests (hematology, coagulation, serum chemistry [includingliver function tests and glucose test], and urinalysis) will beperformed at screening; on Day 1 and Day 85 before the start of infusionand at 1.25 (end of infusion, ±5 minutes), 8 hours (±5 minutes) afterthe start of the infusion; prior to discharge, and all other studyvisits. ^(i) A serum pregnancy test will be performed for females ofchildbearing potential only at screening and at the end-of-study visit.Urine pregnancy tests will be performed on all other study visits, inDay 1 only to confirm the results, no need to have a test again. ^(j)Female subjects who are postmenopausal will have a serum FSH test atscreening. ^(k) Blood samples for the measurement of totalimmunoglobulin/Allergen-specific immunoglobulin E will be collected atscreening, on Day 1 before the start of infusion); and on all otherstudy visits. ^(l) Blood samples for the measurement of biomarkersincluding thymus and activation regulated chemokine (TARC), Eotaxin-3,thymic stromal lymphopoietin (TSLP), periostin, IL-1 , IL-4, IL-5,IL-13, IL-16, IL-31 and M-CSF will be collected at Day 1 and 85 (in 2hours before the start of infusion), and on Days 8, 15, 29, 57, 92, 99,113, and 141. ^(m)The X in Day 1 of Urine drug and blood alcohol testmeans to confirm the results, no need to have a test again

TABLE 3 Baseline Characteristics FB825 Score 5 mg/Kg q12w + TCS (n = 12)range median (IQR)/n (%) mean Age, years, median — 31.5 (25, 34.5) 30.8(IQR) Male, n (%) — 6 (50) EASI, median (IQR) 0-72 27.4 (17.9, 31) 25.8SCORAD, median (IQR)  0-103 60.5 (48.4, 64.8) 57.9 Patients with IGA =0-4  8 (67) 3.7 4, n (%) SCORAD VAS pruritus 0-10 4.95 (3.95, 6.1) 5.0domain, median (IQR) CORAD VAS sleep 0-10 4.5 (1.0, 5.9) 3.9 domain,median (IQR) % BSA, median (IQR)  0-100 42.5 (27.5, 55.8) 43.8 IgE,median (IQR) — 2828.9 (1534.95, 4029.4) 3379.7

TABLE 4 Overall Summary of Number of Patients With Adverse EventsThrough the 15-168 days Treatment Period-SAF Patients with, n (%): FB8255 mg/Kg q12w + TCS (n = 12) Any TEAE 6 (50) Any drug-related TEAE 3 (25)(possible) Any TEAE causing 0 discontinuation of study drugConjuctivitisa 1 (8) Herpes viral infections 2 (20) Upper respiratory 5(42) infection Rhinorrhea 1 (8) Asthma attack 1 (8) Fever 1 (8) Cough 1(8) prolonged QTC 1 (8) Any death 0 Any TE SAE 0 Any drug-related TE SAE0 Any TE SAE causing 0 discontinuation of study drug Any Severe TEAE 0

Results

This is an open-labeled exploratory study to evaluate safety andefficacy of FB825 in adults with atopic dermatitis (AD). 12 eligiblehuman patients with AD were enrolled in the study and received FB825, 5mg/kg, by 1-hour IV infusion on Day 1 and Day 85. Patients werescheduled to return to the study sites according to the schedule ofevents presented in Table 2 for the safety and efficacy evaluation.

FB825 was Safely Administered to Human Subjects with AD

FB825 was administered to the patients via IV infusion over 1 hour at 5mg/kg on days 1 and 85. There were no deaths and no subject discontinueddue to a treatment emergent adverse event (TEAE) or treatment emergentsevere adverse event (TE SAE). Six subjects developed TEAE, three ofwhich are possibly drug related. Five subjects developed upperrespiratory infection and two subjects contracted Herpes viralinfections. Only one subject was reported to have developed one of thefollowing: conjunctivitis, rhinorrhea, asthma attack, fever, cough orprolonged QTC. (Table 4)

There were no apparent treatment or dose-related trends in clinicallaboratory test results, vital sign measurement, 12-lead ECG results, orphysical examination findings.

Efficacy

Efficacy of FB825 was determined by recording the changes from baselinein Eczema Area and Severity Index (EASI), Investigator Global Assessment(IGA), Severity Scoring of Atopic Dermatitis Index (SCORAD) and PruritusVisual Analogue Scale (VAS) at Days Day 1 and 85 before dosing, Day 2and Day 86 before discharge, and any time of Day 8, 15, 29, 57, 92, 99,113, 141, and 169. Baseline demographic and background variables will besummarized by dose and overall for all subjects in Table 3.

(i) EASI was Reduced Compared to Baseline by FB825

Subjects showed progressively reduced EASI after FB825 administration onDay 1 till approximately Day 55 (approximately 65% reduction). EASIscore started rising after Day 55 until the second FB825 administrationon Day 85 but it is still approximately 40% less compared to baseline.Subjects showed further EASI reduction to approximately 70% reduction onDay 113 and maintained the level till the end of study (EOS). (FIG. 5 ).

(ii) IGA was Reduced Compared to Baseline by FB825

Subjects showed progressively reduced IGA after FB825 administration onDay 1 till Day 55 (approximately 30% reduction). IGA score startedrising after Day 55 until the second FB825 administration on Day 85 butit is still approximately 15% less compared to baseline. Subjects showedfurther IGA reduction to approximately 45% reduction at the lowest onDay 113 and maintained the reduction level of 35% compared to baselinetill the end of study (EOS). (FIG. 6 ).

(iii) SCORAD was Reduced Compared to Baseline by FB825

Subjects showed progressively reduced SCORAD after FB825 administrationon Day 1 till approximately Day 55 (approximately 45% reduction). SCORADscore started rising after Day 55 until the second FB825 administrationon Day 85 but it is still approximately 30% less compared to baseline.Subjects showed further SCORAD reduction to approximately 55% reductionon Day 113 and maintained the level till the end of study (EOS). (FIG. 7).

(iv) VAS was Reduced Compared to Baseline by FB825

Subjects showed progressively reduced VAS after FB825 administration onDay 1 till approximately Day 15 (approximately 45% reduction). VAS scorestarted rising after Day 15 and was maintained at 25% reduction comparedto baseline until the second FB825 administration on Day 85. Subjectsshowed further VAS reduction to approximately 55% reduction on Day 113and maintained the level till the end of study (EOS). (FIG. 8 ).

Other Embodiments

All of the features disclosed in this specification may be combined inany combination. Each feature disclosed in this specification may bereplaced by an alternative feature serving the same, equivalent, orsimilar purpose. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

From the above description, one skilled in the art can easily ascertainthe essential characteristics of the present invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions. Thus, other embodiments are also within the claims.

1-59. (canceled)
 60. A method for treating allergic asthma, the methodcomprising: (i) administering to a subject in need thereof a first doseof an antibody binding to a Cεmx domain of a membrane-bound IgE; and(ii) administering to the subject a second dose of the antibody; whereinthe second dose is administered at least 8 weeks and up to 6 monthsafter the first dose; wherein the antibody comprises a heavy chainvariable region, which comprises (a) a heavy chain complementarydetermining region (CDR) 1, a heavy chain CDR2, and a heavy chain CDR3,and (b) a light chain variable region comprising a light chain CDR1, alight chain CDR2, and a light chain CDR3; and wherein: the heavy chainCDR1 comprises the amino acid sequence of residues 26-36 of SEQ ID NO:9;the heavy chain CDR2 comprises the amino acid sequence of residues 51-66of SEQ ID NO:9, the heavy chain CDR3 comprises the amino acid sequenceof residues 98-106 of SEQ ID NO:9, the light chain CDR1 comprises theamino acid sequence of residues 24-39 of SEQ ID NO:10, the light chainCDR2 comprises the amino acid sequence of residues 55-61 of SEQ IDNO:10; and the light chain CDR3 comprises the amino acid sequence ofresidues 94-102 of SEQ ID NO:10.
 61. The method of claim 60, wherein thesecond dose is administered at least 3 months after the first dose. 62.The method of claim 61, wherein the second dose is administered about 12weeks to about 6 months after the first dose.
 63. The method of claim60, wherein the first dose, the second dose, or both range from 0.5mg/kg to 15 mg/kg.
 64. The method of claim 63, wherein the first dose,the second dose, or both range from 1 mg/kg to 8 mg/kg.
 65. The methodof claim 60, wherein the first dose, the second dose, or both areadministered by intravenous injection.
 66. The method of claim 60,wherein the antibody is a human antibody or a humanized antibody. 67.The method of claim 60, wherein the antibody is a full length antibodyor an antigen-binding fragment thereof.
 68. The method of claim 60,wherein the heavy chain variable region comprises the amino acidsequence of SEQ ID NO: 2, SEQ ID NO:8, or SEQ ID NO:9; and the lightchain variable region comprises the amino acid sequence of SEQ ID NO: 3or SEQ ID NO:10.
 69. The method of claim 68, wherein the heavy chainvariable region comprises the amino acid sequence of SEQ ID NO:9 and thelight chain variable region comprises the amino acid sequence of SEQ IDNO:10.
 70. The method of claim 60, wherein the antibody is an IgG1molecule.
 71. The method of claim 60, wherein the antibody comprises aheavy chain comprising the amino acid sequence of residues 20-466 of SEQID NO:11 and a light chain comprising the amino acid sequence ofresidues 21-239 of SEQ ID NO:12.
 72. The method of claim 71, wherein theantibody comprises a heavy chain comprising the amino acid sequence ofSEQ ID NO:11 and a light chain comprising the amino acid sequence of SEQID NO:12.
 73. The method of claim 60, wherein the subject is a humanpatient having allergic asthma.
 74. The method of claim 60, wherein theantibody is formulated in a pharmaceutical composition comprising theantibody, a buffer, a salt, and a nonionic surfactant.
 75. The method ofclaim 74, wherein the pharmaceutical composition is an aqueous solutionhaving a pH of 5 to
 8. 76. The method of claim 75, wherein the buffer isa histidine buffer, the salt is sodium chloride, and/or the nonionicsurfactant is polysorbate
 80. 77. The method of claim 76, wherein theantibody in the pharmaceutical composition is about 10 mg/ml to 30mg/ml, the histidine buffer is of a concentration of about 10 mM to 30mM, the sodium chloride is of a concentration of about 120 mM to 160 mM,and the polysorbate 80 is of a concentration of about 0.01% to 0.03%.